Pharmaceutical compositions

ABSTRACT

The present invention relates to the field of methods for providing pharmaceutical compositions comprising poorly water-soluble drugs. In particular the present invention relates to compositions comprising stable, amorphous hybrid nanoparticles, comprising at least one protein kinase inhibitor and at least one polymeric stabilizing and matrix-forming component, useful in pharmaceutical compositions and in therapy.

This application is a continuation of U.S. patent application Ser. No.16/173,466, filed on Oct. 29, 2018, which is a continuation of U.S.patent application Ser. No. 15/791,093, filed on Oct. 23, 2017, now U.S.Pat. No. 10,143,683, which is a continuation of U.S. patent applicationSer. No. 15/248,107, filed on Aug. 26, 2016, now U.S. Pat. No.9,827,230, which is a continuation of U.S. patent application Ser. No.14/371,875, filed on Jul. 11, 2014, now U.S. Pat. No. 9,456,992, whichis a U.S. National Stage Application of PCT/SE2013/050016, filed on Jan.11, 2013. Priority is claimed to Swedish Patent Application Nos.1250015-3, filed on Jan. 13, 2012 and 1251160-6, filed on Oct. 12, 2012.Priority is also claimed to U.S. Provisional Patent Application Nos.61/586,187, filed on Jan. 13, 2012 and 61/713,120, filed on Oct. 12,2012. The subject matter disclosed in each of the above-mentionedapplications is incorporated herein by reference.

FIELD OF THE INVENTION

The present invention relates to the field of pharmaceuticalcompositions comprising poorly water-soluble drugs. In particular thepresent invention relates to pharmaceutical compositions comprisingstable, amorphous hybrid nanoparticles of protein kinase inhibitors(PKIs) and polymeric stabilizing and matrix-forming components.Furthermore, the present invention relates to a method of treatingproliferative disorders in a patient in need thereof, comprisingadministering a therapeutically effective amount of said compositions.

BACKGROUND OF THE AND INVENTION

Components of cellular signal transduction pathways that regulate thegrowth and differentiation of normal cells can, when dysregulated, leadto the development of cellular proliferative disorders and cancer.Mutations in cellular signaling proteins may cause such proteins tobecome expressed or activated at inappropriate levels or atinappropriate times during the cell cycle, which in turn may lead touncontrolled cellular growth or changes in cell-cell attachmentproperties.

Many proliferative disorders, such as tumors and cancers, have beenshown to involve overexpression or upregulation of protein kinaseactivity.

Protein kinases are kinase enzymes that modify proteins by chemicallyadding phosphate groups (phosphorylation). Phosphorylation usuallyresults in a functional change of the target protein by changing enzymeactivity, cellular location, or association with other proteins. Proteinkinases can be subdivided or characterised by the amino acids of thetarget protein whose phosphorylation they control: most kinases act onboth serine and threonine, the tyrosine kinases act on tyrosine, and anumber (dual-specificity kinases) act on all three. There are alsoprotein kinases that phosphorylate other amino acids, includinghistidine kinases that phosphorylate histidine residues. The humangenome contains about 500 protein kinase genes and up to 30% of allhuman proteins may be modified by protein kinases. Kinases are known toregulate the majority of cellular pathways, especially those involved insignal transduction. Dysregulation of protein kinases by mutation, generearrangement, gene amplification, and overexpression of both receptorand ligand has been implicated in the development and progression ofhuman cancers. Protein kinase inhibiting compounds or protein kinaseinhibitors (PKIs) are therefore useful for treating diseases caused byor exacerbated by overexpression or upregulation of protein kinases. Forexample, tyrosine kinase inhibitors (TKIs also known as tyrphostins)have been shown be effective anti-tumor agents and anti-leukemic agents(Lowery A et. al., Front Biosci. 2011 Jun. 1; 17:1996-2007).

A major objective of formulation chemistry is to improve drug efficiencyand safety, by e.g. improving bioavailability and stability as well asconvenience to the patient. Bioavailability means the rate and extent towhich an active substance or therapeutic is absorbed from apharmaceutical form and becomes available at the site of action. Themost common and preferred method of delivery due to convenience, ease ofingestion, and high patient compliance to treatment is the oral route ofdrug delivery. However, for certain drugs, drug absorption from thegastrointestinal tract is limited by poor aqueous solubility and/or poormembrane permeability of the drug molecules.

PKIs are generally weak bases that dissolve only slightly at low pH(e.g. 100-1000 mg/L) and are practically insoluble at neutral pH (e.g.0.1-10 mg/L). Therefore, enhancing the solubility and dissolution rateof PKI-based drugs is important for improving the bioavailabitity andefficacy of most of these drugs. Typical PKIs exhibit non-polypetidestructure and have relatively low molecular weights, such as ≤10000dalton or ≤5000 dalton.

Several methods to improve the dissolution characteristics of poorlywater soluble drugs have been reported, including micronisation,formation of salts or solvates, complexes and microspheres.Additionally, attempts have been made to improve bioavailabilityprovided by solid dosage forms by forming particles comprising the drugor by mixing the poorly water soluble drug with hydrophilic excipients.Traditionally, however, these methods carry inherent limitationsconcerning physical stabilities of the particles on storage, problemswith grinding or difficulty of removal of the frequently toxic solvent.Furthermore, it is important that the drug released from the solid phasedoes not precitipitate in the gastrointestinal tract, or precipitates aslittle as possible, but remains water-soluble in the aqueous fluids ofthe gastrointestinal tract, since such precipitation results in lowbioavailability (see e.g. Nerve J. et al. Pharm Dev Technol. 2011 June;16(3):278-86).

pH-dependent solubility is a well-known issue for many oral formulationsof poorly water-soluble substances, such as PKIs, since most of theabsorption of the drug occurs in the small and large intestine, where pHis close to neutral. There is thus a continuing need to develop andimprove the dissolution characteristics of oral solid dosage forms ofPKI-based drugs. (Budha N R, Frymoyer A, Smelick G S, Jin J Y, Yago M R,Dresser M J, Holden S N, Benet L Z, Ware J A. Clin Pharmacol Ther. 2012August; 92(2):203-13). Therefore, methods for improving dissolution ofPKI-based drugs, as well as of other poorly water-soluble drugs, atneutral (intestinal) pH are highly desirable.

US20090203709 discloses a pharmaceutical dosage form comprising a soliddispersion product of at least one tyrosine kinase inhibitor, at leastone pharmaceutically acceptable polymer and at least onepharmaceutically acceptable solubilizer. Further the reference disclosesmethods for preparing the above-mentioned pharmaceutical dosage form,comprising preparing the homogenous melt of at least one tyrosine kinaseinhibitor, at least one pharmaceutically acceptable polymer and at leastone pharmaceutically acceptable solubilizer, and allowing the melt tosolidify to obtain a solid dispersion product.

EP2105130 discloses pharmaceutical formulations comprising a soliddispersion or solid solution, containing a polymer and an active agentin amorphous form. Further, the formulation comprises an externalpolymer to stabilize the solution, such that the % by weight of theexternal polymer is less than 20% of the total weight of thepharmaceutical formulation. Additionally, the reference discloses a hotmelt extrusion method for production of the above-mentioned formulation.

SUMMARY OF THE INVENTION

The present invention relates pharmaceutical compositions comprisingstable, amorphous hybrid nanoparticles, comprising at least one proteinkinase inhibitor and at least one polymeric stabilizing andmatrix-forming component. Optionally, one or more solubilizers may beadded to the particles, present separately from the particles, or withinthe particles.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 provides a graph showing the apparent solubility for nilotinib inrepresentative compositions of the invention. Further experimentationwith both nilotinib base and nilotinib HCl is found in Example 1. Thedetails of the particles are described in Example 1, Table 1, forexperiment 3, 30 and 37, respectively. Briefly, experiment 30 representsstable, amorphous hybrid nanoparticles comprising nilotinib HCl andHPMCP HP55 and wherein the solubilizer polyvinyl caprolactam-polyvinylacetate-polyethylene glycol copolymer is present separately from thehybrid nanoparticles. Experiment 3 represents raw, crystalline nilotinibHCl and experiment 37 represents hybrid nanoparticles of nilotinib HCl,HPMCP HP55 and the solubilizer polyvinyl caprolactam-polyvinylacetate-polyethylene glycol copolymer, present within the hybridnanoparticles. The experiments illustrated in the graphs were carriedout at pH 6.5, in FaSSIF.

FIG. 2 provides a graph showing the apparent solubility for erlotinib inrepresentative compositions of the invention. Further experimentationwith erlotinib is found in Example 2. The details of the stable,amorphous hybrid nanoparticles are described in Example 2, Table 7, forexperiment 58, 65 and 67, respectively. Briefly, experiment 65represents stable, amorphous hybrid nanoparticles with erlotinib HCl andHPMC-AS, wherein the solubilizer polyvinyl caprolactam-polyvinylacetate-polyethylene glycol copolymer is present separately from thestable, amorphous hybrid nanoparticles. Experiment 58 represents raw,crystalline erlotinib HCl and experiment 67 represents stable, amorphoushybrid nanoparticles of erlotinib HCl, HPMC-AS and the solubilizerpolyvinyl caprolactam-polyvinyl acetate-polyethylene glycol copolymerpresent within the hybrid nanoparticles. The experiments illustrated inthe graphs were carried out at pH 6.5 in FaSSIF.

FIG. 3 provides a graph showing the apparent solubility for pazopanib inrepresentative compositions of the invention. Further experimentationwith pazopanib is found in Example 3. The details of the stable,amorphous hybrid nanoparticles are described in Example 3, Table 13, forexperiment 84, 91 and 93, respectively. Briefly, experiment 91represents stable, amorphous hybrid nanoparticles comprising pazopaniband PVP 90K and wherein the solubilizer polyvinyl caprolactam-polyvinylacetate-polyethylene glycol copolymer is present separately from thestable, amorphous hybrid nanoparticles, experiment 93 represents hybridnanoparticles comprising pazopanib, PVP 90K and the solubilizerpolyvinyl caprolactam-polyvinyl acetate-polyethylene glycol copolymer,present within the stable, amorphous hybrid nanoparticles. Experiment 84represents raw, crystalline pazopanib. The experiments illustrated inthe graphs were carried out at pH 6.5, in FaSSIF.

FIG. 4 provides a graph showing the apparent solubility for lapatinib inrepresentative compositions of the invention. Further experimentationwith both lapatinib base and lapatinib ditosylate salt is found inExample 4. The details of the stable, amorphous hybrid nanoparticles aredescribed in Example 4, Table 19, for experiment 110, 122 and 126,respectively. Briefly, experiment 122 represents stable, amorphoushybrid nanoparticles comprising lapatinib base and HPC EF, wherein thesolubilizer polyvinyl caprolactam-polyvinyl acetate-polyethylene glycolcopolymer is present separately from the stable, amorphous hybridnanoparticles. Experiment 110 represents raw, lapatinib base andexperiment 126 represents stable, amorphous hybrid nanoparticles oflapatinib base, HPC LF and the solubilizer polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol copolymer presentwithin the hybrid nanoparticles. The experiments illustrated in thegraphs were carried out at pH 6.5 in FaSSIF.

FIG. 5 provides a graph showing the apparent solubility for nilotinib inrepresentative compositions of the invention. The details of the stable,amorphous hybrid nanoparticles are described in Example 5, Table 21, forexperiment 127, 128 and 129, respectively. Briefly, experiment 129represents a physical mixture of raw, crystalline nilotinib HCl, HPMCPHP55 and the solubilizer polyvinyl caprolactam-polyvinylacetate-polyethylene glycol copolymer. Experiment 128 represents stable,amorphous hybrid nanoparticles comprising nilotinib HCl and HPMCP HP55,wherein the solubilizer polyvinyl caprolactam-polyvinylacetate-polyethylene glycol copolymer is present separately from thestable, amorphous hybrid nanoparticles. Experiment 127 representsstable, amorphous hybrid nanoparticles of nilotinib HCl and HPMCP HP55.The experiments illustrated in the graphs were carried out at pH 1.4 inSGF.

FIG. 6 provides a graph showing the apparent solubility for gefitinib inrepresentative compositions of the invention. Further experimentationwith gefitinib is found in Example 6. The details of the compositionsare described in Example 6, Table 22, for experiment 131, 133, 135 and137, respectively. Briefly, experiment 131 represents raw, crystallinegefitinib. Experiment 133 represents a mixture of raw, crystallinegefitinib, HPMCP HP55 and the solubilizer polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol copolymer. Experiment135 represents stable, amorphous hybrid nanoparticles of gefitinib andHPMCP HP55. Experiment 137 represents stable, amorphous hybridnanoparticles of gefitinib and HPMCP HP55 wherein the solubilizerpolyvinyl caprolactam-polyvinyl acetate-polyethylene glycol copolymer ispresent separately from the stable, amorphous hybrid nanoparticles. Theexperiments illustrated in the graphs were carried out at pH 6.5 inFaSSIF.

FIG. 7 provides a graph showing the apparent solubility for dasatinib inrepresentative compositions of the invention. The details of the stable,amorphous hybrid nanoparticles are described in Example 7, Table 24, forexperiments 138-141. Briefly, experiment 138 represents raw, crystallinedasatinib. Experiment 139 represents a mixture of raw, crystallinedasatinib, Kollidon VA64 and the solubilizer polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol copolymer. Experiment140 represents hybrid nanoparticles of dasatinib and Kollidon VA64.Experiment 141 represents stable, amorphous hybrid nanoparticles ofdasatinib and Kollidon VA64 wherein the solubilizer polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol copolymer is presentseparately from the stable, amorphous hybrid nanoparticles. Theexperiments illustrated in the graphs were carried out at pH 6.5 inFaSSIF.

FIG. 8 provides a graph showing the apparent solubility for sorafenib inrepresentative compositions of the invention. The details of the stable,amorphous hybrid nanoparticles are described in Example 8, Table 26, forexperiments 142-145. Briefly, experiment 142 represents raw, crystallinesorafenib tosylate. Experiment 143 represents a mixture of raw,crystalline sorafenib tosylate, HPMCP HP55 and the solubilizer polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol copolymer. Experiment144 represents stable, amorphous hybrid nanoparticles of sorafenibtosylate and HPMCP HP55. Experiment 145 represents hybrid nanoparticlesof sorafenib tosylate and HPMCP HP55 wherein the solubilizer polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol copolymer is presentseparately from the stable, amorphous hybrid nanoparticles. Theexperiments illustrated in the graphs were carried out at pH 6.5 inFaSSIF.

FIG. 9 provides a graph showing the apparent solubility for crizotinibin representative compositions of the invention. Further experimentationwith crizotinib is found in Example 10. The details of the compositionsare described in Example 10, Table 30, for experiment 150, 152, 153 and156, respectively. Briefly, experiment 150 represents raw, crystallinecrizotinib. Experiment 152 represents a mixture of raw, crystallinecrizotinib, PVP 30K and the solubilizer Cremophor RH40. Experiment 153represents stable, amorphous hybrid nanoparticles of crizotinib and PVP30K. Experiment 156 represents stable, amorphous hybrid nanoparticles ofcrizotinib and PVP 30K wherein the solubilizer Cremophor RH40 is presentseparately from the stable, amorphous hybrid nanoparticles. Theexperiments illustrated in the graphs were carried out at pH 6.5 inFaSSIF.

FIG. 10 provides a graph showing the apparent solubility for axitinib inrepresentative compositions of the invention. Further experimentationwith axitinib is found in Example 11. The details of the compositionsare described in Example 11, Table 32, for experiment 157, 158, 160 and162, respectively. Briefly, experiment 157 represents raw, crystallineaxitinib. Experiment 158 represents a mixture of raw, crystallineaxitinib, Kollidon VA64 and the solubilizer polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol copolymer. Experiment160 represents stable, amorphous hybrid nanoparticles of axitinib andKollidon VA64. Experiment 162 represents stable, amorphous hybridnanoparticles of axitinib and Kollidon VA64 wherein the solubilizerpolyvinyl caprolactam-polyvinyl acetate-polyethylene glycol copolymer ispresent separately from the stable, amorphous hybrid nanoparticles. Theexperiments illustrated in the graphs were carried out at pH 6.5 inFaSSIF.

FIG. 11 provides a graph showing the apparent solubility for vemurafenibin representative compositions of the invention. Further experimentationwith vemurafenib is found in Example 12. The details of the compositionsare described in Example 12, Table 34, for experiment 164, 166, 168 and170, respectively. Briefly, experiment 164 represents raw, crystallinevemurafenib. Experiment 166 represents a mixture of raw, crystallinevemurafenib, CAP and the solubilizer polyvinyl caprolactam-polyvinylacetate-polyethylene glycol copolymer. Experiment 168 represents stable,amorphous hybrid nanoparticles of vemurafenib and CAP. Experiment 170represents stable, amorphous hybrid nanoparticles of vemurafenib and CAPwherein the solubilizer polyvinyl caprolactam-polyvinylacetate-polyethylene glycol copolymer is present separately from thestable, amorphous hybrid nanoparticles. The experiments illustrated inthe graphs were carried out at pH 6.5 in FaSSIF.

FIG. 12 provides a graph showing the dissolution rate for nilotinib basein representative compositions of the invention, measured under sinkconditions. Details are found in Examples 13 and 13.1, and Table 36 forexperiments 500 and 501. Briefly, experiment 500 represents raw,nilotinib HCl. Experiment 501 represents stable, amorphous hybridnanoparticles of nilotinib base and HPMCP HP55. The experimentsillustrated in the graphs were carried out at pH 6.5 in FaSSIF.

FIG. 13 provides a graph showing the dissolution rate for erlotinib inrepresentative compositions of the invention, measured under sinkconditions. Details are found in Examples 13 and 13.2, Table 37 forexperiments 510 and 511. Briefly, experiment 510 represents raw,erlotinib HCl. Experiment 511 represents stable, amorphous hybridnanoparticles of erlotinib HCl and HPMC AS.

FIG. 14 provides a graph showing the dissolution rate for pazopanib inrepresentative compositions of the invention, measured under sinkconditions. Details are found in Examples 13 and 13.3, Table 38 forexperiments 520 and 521. Briefly, experiment 520 represents raw,pazopanib HCl. Experiment 521 represents stable, amorphous hybridnanoparticles of pazopanib HCl and PVP90K.

FIG. 15 provides a graph showing the dissolution rate for lapatinib inrepresentative compositions of the invention, measured under sinkconditions. Details are found in Examples 13 and 13.4, Table 39 forexperiments 530 and 531. Briefly, experiment 530 represents raw,lapatinib ditosylate. Experiment 531 represents stable, amorphous hybridnanoparticles of lapatinib base and HPC lf.

FIG. 16 provides a graph showing the dissolution rate for gefitinib inrepresentative compositions of the invention, measured under sinkconditions. Details are found in Examples 13 and 13.5. Table 40 forexperiments 540 and 541. Briefly, experiment 540 represents raw,gefitinib. Experiment 541 represents stable, amorphous hybridnanoparticles of gefitinib and HPMCP HP55.

FIG. 17 provides a graph showing the dissolution rate for dasatinib inrepresentative compositions of the invention, measured under sinkconditions. Details are found in Examples 13 and 13.6. Table 41 forexperiments 550 and 551. Briefly, experiment 550 represents raw,dasatinib. Experiment 551 represents stable, amorphous hybridnanoparticles of dasatinib and Kollidon VA64.

FIG. 18 provides a graph showing the dissolution rate for sorafenib inrepresentative compositions of the invention, measured under sinkconditions. Details are found in Examples 13 and 13.7. Table 42 forexperiments 560 and 561. Briefly, experiment 560 represents raw,sorafenib tosylate. Experiment 561 represents stable, amorphous hybridnanoparticles of sorafenib tosylate and HPMCP HP55.

FIG. 19 provides a graph showing the dissolution rate for crizotinib inrepresentative compositions of the invention, measured under sinkconditions. Details are found in Examples 13 and 13.8. Table 43 forexperiments 570 and 571. Briefly, experiment 570 represents raw,crizotinib. Experiment 571 represents stable, amorphous hybridnanoparticles of crizotinib and PVP 30K.

FIG. 20 provides a graph showing the dissolution rate for axitinib inrepresentative compositions of the invention, measured under sinkconditions. Details are found in Examples 13 and 13. 9. Table 44 forexperiments 580, 581 and 582. Briefly, experiment 580 represents raw,axitinib. Experiment 581 represents hybrid nanoparticles of axitinib andKollidon VA64 and experiment 582 represents stable, amorphous hybridnanoparticles of axitinib and HPMC AS.

FIG. 21 provides a graph showing the dissolution rate for vemurafenib inrepresentative compositions of the invention, measured under sinkconditions. Details are found in Examples 13 and 13.10. Table 45 forexperiments 590, 591 and 592. Briefly, experiment 590 represents raw,vemurafenib. Experiment 591 represents hybrid nanoparticles ofvemurafenib and Kollidon VA64 and experiment 592 represents stable,amorphous hybrid nanoparticles of vemurafenib and CAP.

FIG. 22 provides graphs showing in vivo measurement of plasma levelsafter oral administration to beagle dogs of representative compositionscomprising stable, amorphous hybrid nanoparticles of nilotinib base andthe polymeric stabilizing and matrix-forming components PVAP and HPMCPHP55, respectively (UP), denoted PVAP and HP55, as well as wherein thesolubilizer polyvinyl caprolactam-polyvinyl acetate-polyethylene glycolcopolymer was added (I/P+S), denoted HP55s and PVAPs, respectively. Theexperiments were carried out in beagle dogs pre-treated to have neutralstomach content. The stable, amorphous hybrid nanoparticles are furtherdescribed in experiments 146 and 147 (Example 9) and details of the invivo experiments are set out in Example 14. The experiments used amarketed formulation comprising nilotinib HCl (“Tasigna”) as reference.

FIG. 23 provides graphs showing in vivo measurement of plasma levelsafter oral administration to beagle dogs of representative compositionscomprising stable, amorphous hybrid nanoparticles of nilotinib base andthe polymeric stabilizing and matrix-forming components PVAP and HPMCPHP55, respectively (UP), denoted PVAP and HP55, as well as wherein thesolubilizer polyvinyl caprolactam-polyvinyl acetate-polyethylene glycolcopolymer was added (I/P+S) denoted PVAPs and HP55s, respectively. Theexperiments were carried out in beagle dogs pre-treated to have acidicstomach content. The stable, amorphous hybrid nanoparticles are furtherdescribed in experiments 146 and 147 (Example 9) and details of the invivo experiments are set out in Example 14. The experiments used amarketed formulation comprising nilotinib HCl (“Tasigna”) as reference.

FIG. 24 provides graphs showing in vivo measurement of plasma levelsafter oral administration to beagle dogs of representative compositionscomprising stable, amorphous hybrid nanoparticles of nilotinib base andthe polymeric stabilizing and matrix-forming components PVAP and HPMCPHP55, respectively (I/P), denoted PVAP and HP55, as well as wherein thesolubilizer polyvinyl caprolactam-polyvinyl acetate-polyethylene glycolcopolymer was added after hybrid nanoparticle formation (I/P+S), denotedPVAPs and HP55s, respectively. The experiments were carried out inbeagle dogs pre-treated to have acidic or neutral stomach content. Thestable, amorphous hybrid nanoparticles are further described inexperiments 146 and 147 (Example 9) and details of the in vivoexperiments are set out in Example 14.

FIG. 25 provides graphs showing in vivo measurement of plasma levelsafter oral administration to beagle dogs of representative compositionscomprising stable, amorphous hybrid nanoparticles of nilotinib base andthe polymeric stabilizing and matrix-forming components PVAP and HPMCPHP55, denoted PVAP and HP55, respectively (I/P). The experiments werecarried out in beagle dogs pre-treated to have acidic or neutral stomachcontent. The stable, amorphous hybrid nanoparticles are furtherdescribed in experiments 146 and 147 (Example 9) and details of the invivo experiments are set out in Example 14.

FIG. 26 provides a graph showing the apparent solubility ofrepresentative compositions before and after 11 months of storage atroom temperature. The experiment provides stable, amorphous hybridnanoparticles comprising nilotinib base, HPMCP HP55 and the addition ofthe solubilizer polyvinyl caprolactam-polyvinyl acetate-polyethyleneglycol copolymer (I/P+S) as Exp 171 & Exp 172 with further details setout in Example 15.

FIG. 27 provides overlayed X-ray powder diffraction (XRPD) patterns ofstable hybrid nanoparticles at 40% drug load, I/P nilotinib base/HPMCPHP55. Initial (top) and after 12 months storage at ambient temperature(bottom). The XRPD patterns are offset in order improve the visualcomparison. Further details are set out in Example 15.

DETAILED DESCRIPTION OF THE INVENTION

All patents, patent applications, and publications cited herein arehereby incorporated by reference in their entirety.

As used herein, the phrase “hybrid nanoparticles” refers to a group ofparticles, typically in the average size range of from 1 to 1000 nm,composed of at least two components, one of which is the PKI and theother a polymeric stabilizing and matrix-forming component. Theparticles can be either crystalline or amorphous, or a mixture thereof.Typically, in the sense of the present disclosure, the particles are“amorphous”, or “essentially amorphous”. This means that almost all, ifnot all, content of the particles comprise amorphous protein kinaseinhibitor and polymeric stabilizing and matrix-forming component. Thelevel or degree of amorphicity is at least 60%, such as 70%, such as 80%or 85%, preferably at least 90% and more preferably >95%, wherein 100%represents that all material is amorphous in the particles.

Quantification of crystalline PKI or absence of crysalline PKI may bemeasured by X-ray powder diffraction metods as described inSaleki-Gerhardt A et al. Int J Pharm. 1994; 101:237-247) or by watervapor sorption as described in Dash A K et al. J Pharm Sci. 2002 April;91(4):983-90.

The term “solid dispersion particles” relates to “hybrid nanoparticles”as defined above, however, solid dispersion particles are typicallylarger or much larger in size (typically μm-mm, as described in Wu K. etal. J Pharm Sci. 2009 July; 98(7):2422-3). The smaller size of hybridnanoparticles contributes to further stabilizing the PKI againstcrystallization. Typically, hybrid nanoparticles is in the average sizerange of from 1 to 1000 nm, such as below 500 nm, preferably below 250nm.

The phrase “stable” refers to the level of stability of producedparticles by the methods of the present invention and may be measured asthe capability of the hybrid nanoparticles to remain in their physicalstate for 6-12 months storage at ambient temperature (e.g. 18-25° C.).The level of stability may be measured by AUC measurements ofdissolution rate over for instance 80 minutes of the particles, aftersuch storage.

By the phrase “protein kinase inhibitor” or “PKI” is meant a type ofenzyme inhibitor that specifically blocks the action of one or moreprotein kinases. PKIs include, but are not limited, to protein kinaseinhibitors and tyrosine kinase inhibitors, such as axitinib, afatinib,bosutinib, crizotinib, cediranib, dasatinib, erlotinib, fostamatinib,gefitinib, imatinib, lapatinib, lenvatinib, lestaurtinib, motesanib,mubritinib, nilotinib, pazopanib, pegaptanib, ruxolitinib, sorafenib,semaxanib, sunitinib, tandunitib, tipifamib, vandetanib and vemurafenib;or salts or hydrates or solvates thereof, or combinations thereof.

By the phrase “polymeric stabilizing and matrix-forming component” ismeant the component present in the hybrid nanoparticles together withthe PKI. Typically, said polymeric stabilizing and matrix-formingcomponent exhibits a polymeric structure, such as, but not limited to,methyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose (e.g.HPC ef, HPC lf and HPC jf), hydroxypropyl methylcellulose (e.g. MethocelE3 and E15 and Pharmacoat), hydroxypropyl methylcellulose acetatesuccinate (HPMC AS), hydroxypropyl methylcellulose phthalate (e.g.HPMCP-HP55), polyvinylpyrrolidone (e.g. PVP 30K and PVP 90K), polyvinylacetate phthalate (PVAP), copolyvidone (e.g. Kollidon VA 64),crospovidon (e.g. Kollidon CL), methacrylic acid and ethylacrylatecopolymer (e.g. Kollicoat ME), methacrylate acid and methyl methacrylatecopolymer (e.g. Eudragit L100), polyethylene glycol (PEG), DLlactide/glycolide copolymer, poly DL-lactide, cellulose acetatephthalate (CAP), aminoalkyl methacrylate copolymers (e.g. EudragitRL100, RL PO or RS PO), carbomer homopolymer Type A (e.g. Carbopol971P), carbomer homopolymer Type B (e.g. Carbopol 974P) and Poloxamers(e.g. Pluronics, Kolliphor).

The term “polymer” or “polymeric” is here used to mean a compound thatis made of monomers connected together to form a larger molecule. Apolymer generally consists of 20 or more monomers connected together,however less than 20 monomers connected together are here also referredto as polymers.

The term “solubilizer” is here used to mean a compound that increasesthe solubility of a substance, such as, but not limited to, polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol copolymer (Soluplus),d-α-tocopherol acid polyethylene glycol 1000 succinate (TPGS), PEG-40hydrogenated castor oil (Cremophor RH40), PEG-35 castor oil (CremophorEL), PEG-40 stearate (MYRJ 540), hard fat (e.g. Gelucire 33/01),polyoxylglycerides (e.g. Gelucire 44/14), stearoyl polyoxylglycerides(e.g. Gelucire 50/13), PEG-8 caprylic/capric glycerides (e.g. Labrasol)and Poloxamers (e.g. Pluronics, Kolliphor).

As used herein, the phrase “primary particles” refers to the smallestparticulate entities formed during the precipitation process. Theboundaries of the particles are analyzed by SEM microscopy. Depending onprocess parameters, the primary particles may build together a more orless dense and porous network forming larger, agglomerated or bridgingparticles. Parameters affecting the agglomeration are e.g. temperaturethat may modify the softness of the primary particles; ratiosolvent/antisolvent affecting precipitation time, concentration of thePKI solution; and the nature of the polymeric stabilizing andmatrix-forming agent(s). The average size of the primary particles istypically between 1 to 1000 nm, preferably below 500 nm, more preferablybelow 250 nm.

As used herein, the phrases “supercritical” and “supercritical fluid”refer to that a chemical substance that is set to both a temperaturehigher or equal than its critical temperature (Tc) and a pressure higheror equal than its critical pressure (Pc).

As used herein, the phrases “subcritical” and “subcritical fluid” referhere to that one of critical temperature (Tc) or critical pressure (Pc)is set to a temperature or pressure higher than its critical temperature(Tc) or critical pressure (Pc), respectively, and the other of criticaltemperature (Tc) or critical pressure (Pc) is set to a temperature orpressure lower than its critical temperature (Tc) or critical pressure(Pc), respectively.

By the phrase “area under the curve (AUC)” is meant the area under theconcentration-time curve, where the x-axis represents time and they-axis represents solubilized drug concentration.

By the phrase “apparent solubility” is meant the concentration ofmaterial at apparent equilibrium. See further in the Examples section.

The term “supersaturation” is here used to mean that a solution containsmore of the dissolved substance than could be dissolved by the solventor media under normal circumstances.

As used herein, the term “Soluplus” or “soluplus” refers to polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol copolymer.

As used herein, the term “TPGS” refers to d-α-tocopherol acidpolyethylene glycol 1000 succinate.

As used herein, the term “Chremophor RH40” refers to PEG-40 hydrogenatedcastor oil.

As used herein, the term “PVAP” refers to polyvinyl acetate phthalate.

As used herein, the term “PVP 90K” refers to polyvinylpyrrolidone K-90.

As used herein, the term “PVP 30K” refers to polyvinylpyrrolidone K-30.

As used herein, the term “HPMC-AS” refers to hydroxypropylmethylcellulose acetate succinate.

As used herein, the term “HPMCP HP55” refers to hydroxypropyl methylcellulose phthalate.

As used herein, the term “HPC” refers to hydroxypropyl cellulose, suchas HPC EF and HPC LF.

As used herein, the term “Kollidon VA64” refers to copolyvidone.

As used herein, the term “CAP” refers to cellulose acetate phthalate.

The dissolution mediums used for purposes of testing hybridnanoparticles of the present invention, includes Fasted State StimulatedIntestinal Fluid, referred to as FaSSIF, Fed State Stimulated IntestinalFluid, referred to as FeSSIF, and Simulated Gastric Fluid, referred toas SGF. FaSSIF media is tailored to represent a fasting state and has apH of about 6.5 as well as particular osmolaric properties. FeSSIF mediais tailored to represent a fed state and has a pH of about 5 as well asspecific osmolaric properties. SGF is tailored to represent gastricfluid and has a pH of about 1.4 as well as particular osmolaricproperties. FaSSIF, FeSSIF and SGF media are generally used in in vitromodels for dissolution of poorly water-soluble drugs. The choice ofmedium will be dependent of the where in the intestinal tract and underwhat conditions (fasted or fed) particles are desired to dissolve and betaken up. Further details regarding these fluids are described in e.g.Hervé J. et al. Pharm Dev Technol. 2011 June; 16(3):278-86 andJantratid, E., and Dressman, J. Dissolut. Technol. 2009 8, 21-25.

By the phrase “amorphous form” is meant non-crystalline solid form. Theease of dissolution may at least in part be attributed to the amount ofenergy required for dissolution of the components from a crystalline oramorphous solid phase. Amorphous particles require less energy fordissolution as compared to crystalline particles of the same compound.

The inventive compositions comprise particles with a PKI or acombination of two or more PKIs. However, the particles may comprise acombination of one or more PKIs and at least one further activeingredient, such as one or more drugs. Various kinds of PKIs can beeffectively utilized.

The term PKIs (protein kinase inhibitors) as used herein, is intended toinclude also the hydrates, solvates (alcoholates), pharmaceuticallyacceptable acid salts, base salts or co-crystals of such protein kinaseinhibiting compounds.

As used herein, the term water-insoluble or poorly water soluble (orhydrophobic) compounds, refers to compounds whose solubility in water at25° C. is less than 1 g/100 ml, especially less than 0.1 g/100 ml inpure water at neutral pH.

The stable, amorphous hybrid nanoparticles comprised in the compositionsof the present invention are typically in the form of particles asdescribed elsewhere in this specification. There are a number ofdifferent methods for the formation of larger particles, e.g.granulation, melt extrusion, spray drying, precipitation etc. all ofwhich typically encompass starting with formation of a mixture betweenthe Active Pharmaceutical Ingredient (API) and the polymeric stabilizingand matrix-forming component. The particles comprised in thecompositions of the present invention are produced with continuousprocesses for generating hybrid nanoparticles. Continuous processes inthis context means that particle formation is continuously ongoing whileat the same time continuously withdrawing/collecting/retaining hybridnanoparticles from the mixture after their formation. In the preferredmethods, i.e. precipitation methods, this means that a fluid which is asolution of the PKI, preferably in the form of a fluid stream, is mixedwith an antisolvent fluid, preferably in the form of an antisolventfluid stream. The polymeric stabilizing and matrix-forming component maybe present in either one or both of the two fluids depending on itssolubility characteristics. The mixing of the two fluids is taking placein a mixing function, e.g. a mixing chamber. In the case the process iscontinuous, i.e. the two fluids are fluid streams, the mixing functiontypically is associated with a particle formation and separationfunction wherein the mixed fluid stream may pass through while retainingthe hybrid nanoparticles. Agents modifying the particle characteristicswithout being incorporated into the particles may be added to either oneor both of the two fluids before the mixing step. The fluids typicallyare conventional liquids or supercritical fluids, where supercriticalfluids also include subcritical fluids (i.e. fluids for which only oneof pressure and temperature is above its supercritical value). Typicalcombinations are, a) conventional (i.e., non-supercritical) liquids forboth the API solution and the antisolvent, b) supercritical solution ofthe API combined with conventional liquid for the antisolvent, c)conventional liquid for the API solution combined with supercriticalfluid for the antisolvent, and d) supercitical fluids for both of thetwo fluids. In certain variants the antisolvent may be omitted. A fluidstream, preferably supercritical, containing both the API and thepolymeric stabilizing and matrix-forming component is then allowed toexpand into the particle formation function. It is preferred that atleast one of the fluids is in a supercritical state in the preciptiationmethods described above. These kinds of precipitation methods arediscussed in WO 2005061090 (Censdelivery AB), WO 2009072950 (XSprayMicroparticles AB), WO 2009072953 (XSpray Microparticles AB), WO2011159218 (XSpray Microparticles AB) and references cited in thesepublications.

The term “solution” encompasses that the solute is either a true soluteor minute particles of colloidial dimensions (typically 1-1000 nm) andless than the particles to be produced.

A preferred particle formation system is the “Right Size system”developed by XSpray Microparticles AB, Sweden. A detailed description ofthe technology can be found in the WO-publications given in thepreceding paragraph. An important characteristic of the system is thatthe two fluid streams should merge within a nozzle at an angle in theinterval 45°-135°, with preference for about 90° and sprayed into aparticle formation/separation function. In principle the system allowsfor producing particles of predetermined size and/or morphology. Herethe Right Size system and apparatus will be described using thenon-limiting example of a PKI as the drug and CO₂ as a supercrititcalfluid antisolvent.

The system consists of one pumping set-up for the PKI dissolved in aliquid solvent, referred to as the API solution, and one pumping set-upfor an antisolvent, for example CO₂, however also other antisolvents maybe used when suitable. Each pumping set-up includes instruments such asa flow meter and a pressure meter that are used to control the processconditions. These two pumping set-ups are fluidically connected at aspray nozzle.

A stream of liquid API solution is mixed with a stream of CO₂ under flowconditions within the spray nozzle. The polymeric stabilizing andmatrix-forming component is present in either the API solution or in thestream of CO₂. These streams are sprayed at the outlet of the nozzleinto a precipitation vessel under controlled conditions (typicallypressure and temperature). CO₂ acts as an antisolvent and makes the APIto precipitate together with the polymeric stabilizing andmatrix-forming component into fine particles. Particles are retained inthe vessel by a filtering set-up. A back pressure regulator is typicallyused to control the pressure inside the precipitation vessel.

For preparing hybrid nanoparticles of certain drugs, for example but notlimited to pazopanib and erlotinib, it may be advantageous to have anextra pumping set-up for injecting an additional solvent, referred to asa modifier, into the CO₂. Here a pumping set-up control is set up forthe modifier and the modifier is mixed with the CO₂ in a mixer beforeentering the nozzle.

When using the system, the system operator typically starts byequilibrating the system by pumping CO₂, an “PKI like solution” (asolution similar in composition to the PKI solution but containing noPKI and no excipient) and the modifier (if used) through the systemuntil flow rates, pressure and temperature have reached a desired steadystate. Critical parameters for setting up the system are PKI solutioncomposition, PKI solution flow rate, CO₂ flow rate, CO₂ pressure andtemperature, nature of the modifier and modifier flow rate, if such isused.

Next, the “PKI like solution” is exchanged for the PKI solution andparticles are produced and retained downstream of the mixing, e.g.downstream of the outlet of the nozzle. Afterwards, the system istypically cleaned by pumping the “PKI like solution” through the system.The particles are dried by flushing CO₂ through the retained particlesin order to extract any remaining solvent. The precipitation vessel isthen depressurized and the particles can be collected.

The solution/solvent and the antisolvent are typically miscible witheach other. The pressure and temperature in the particle formationfunction, and/or upstream of this function, such as in the mixingfunction, provide supercritical or subcritical conditions in relation tothe antisolvent.

The concentration of the PKI in the solution is typically below itssaturation concentration, such as ≤50%, such as ≤60%, such as ≤75%, suchas ≤85% or such as ≤95% of the saturation concentration. Suitableconcentrations are typically found in the interval ≤20%, such as ≤10% or≤5% or ≤3% with lower limits being ≤005% or 0.1% (all in w/v-%). Theterm “volatile” for solvents typically means boiling points of ≤200° C.,such as ≤150° C. or ≤100° C., at atmospheric pressure. Examples areinorganic solvents and organic solvents with particular emphasis ofdimethyl sulfoxide and trifluoroethanol and mixtures thereof. The termsolvent includes mixtures of liquids which are miscible with each other.The solutions may contain agents that enhance or diminish the solubilityof the PKI, e.g. acidic, alkaline, buffer components and/or otherorganic solvents.

Illustrative fluids which can be used as an antisolvent are

a) gaseous at room temperature and atmospheric pressures, or

b) liquid at room temperature and atmospheric pressure.

The antisolvent is typically selected for its ability to be readilydispersed into small droplets and for its ability to act as an atomizingagent and antisolvent against the PKI present in the solution.

Compounds/elements according to group (a) may be selected from carbondioxide (Pc=74 bar and Tc=31° C.) (preferred), nitrous oxide (Pc=72 barand Tc=36° C.), sulphur hexafluoride (Pc=37 bar and Tc=45° C.), ethane(Pc=48 bar and Tc=32° C.), ethylene (Pc=51 bar and Tc=10° C.), xenon(Pc=58 bar and Tc=16° C.), trifluoromethane (Pc=47 bar and Tc=26° C.),chlorotrifluoromethane (Pc=39 bar and Tc=29° C.) and nitrogen (Pc=34 barand Tc=−147° C.) and mixtures containing these compounds/elements. Pcstands for critical pressure and Tc for critical temperature. Compoundsaccording to group (b) are typically selected amongst conventionalliquids of the same general types as discussed for solvents above butwith the difference that the PKI present in the solution must be poorlysoluble in the antisolvent. Particular liquids of group (b) comprisemethanol, ethanol, acetone water and mixtures containing one or more ofthese fluids.

The antisolvents of group (a) above are typically used at pressures andtemperatures providing i) supercritical conditions (supercritical fluid)or ii) a subcritical conditions (subcritical fluid) in the particleformation function and/or upstream of this function, such as in themixing function and upstream of this latter function.

Variant (i) means pressures and temperatures which are typically abovethe critical pressure Pc and critical temperature Tc of the antisolventused. For the pressure this typically means pressures in the interval(1.0-7.0)×Pc or in the interval 10 bar, suitably ≥20 bar with preferencefor ≥30 bar, higher than Pc with illustrative upper limits being 100bar, 200 bar and 300 bar higher than Pc. For the temperature thistypically means temperatures within (1.0-4.0)×Tc or in the interval of≤5° C., suitably ≤10° C. with preference for 15° C. above Tc withillustrative upper limits being 10° C., 40° C. and 50° C. above Tc.

Variant (ii) means that at least one of temperature and pressure, withpreference for only the temperature, is/are below the critical value.(Tc and Pc, respectively). Thus the temperature may be in the intervalof (0.1-1)×Tc, such as (0.5-1)×Tc, or lower. Further, the temperaturemay be low, such as −10° C. or −30° C. These temperatures may becombined with pressures as defined in the preceding paragraph or withpressures lower than the Pc of the used antisolvent. For carbon dioxidethis means that the temperature in the particle formation function is<+31° C., such as about +25° C. or lower combined with a pressure aboveor below 74 bar.

The antisolvents of group (b) above are typically used in thesubcritical state, i.e. as a subcritical fluid.

In one aspect of the invention, there is provided a pharmaceuticalcomposition comprising stable, amorphous hybrid nanoparticles of atleast one protein kinase inhibitor and at least one polymericstabilizing and matrix-forming component; which composition optionallyfurther comprises at least one pharmaceutically acceptable solubilizer.

In one embodiment of this aspect, there is provided a pharmaceuticalcomposition comprising stable, amorphous hybrid nanoparticles of atleast one protein kinase inhibitor and at least one polymericstabilizing and matrix-forming component; which composition furthercomprises at least one pharmaceutically acceptable solubilizer.Typically, said solubilizer is present separated from the hybridnanoparticles in the composition. Or, typically, said solubilizer isdistributed to the surface of the hybrid nanoparticles. Said solubilizermay be selected from polyvinyl caprolactam-polyvinylacetate-polyethylene glycol copolymer, d-α-tocopherol acid polyethyleneglycol 1000 succinate and a hydrogenated castor oil, such as PEG-40hydrogenated castor oil or PEG-35 hydrogenated castor oil. Furthermore,said solubilizer may be a poloxamer. The compositions comprising stable,amorphous hybrid nanoparticles comprising at least one protein kinaseinhibitor and at least one polymeric stabilizing and matrix-formingcomponent, display increased dissolution rate.

Consequently, in another embodiment of this aspect, there is provided acomposition comprising stable, amorphous hybrid nanoparticles,comprising at least one protein kinase inhibitor and at least onepolymeric stabilizing and matrix-forming component, wherein said hybridnanoparticles display an increased dissolution rate of said proteinkinase inhibitor, compared to the dissolution rate of said proteinkinase inhibitor in raw, crystalline form.

Typically, said dissolution rate is measured by a flow through cellsystem in sink conditions, e.g., according to the US Pharmacopea (USP4).Dissolution measurement in sink conditions of hybrid nanoparticles maybe measured in a method consisting of adding the wished amount of powderinto a flow through cell system (SOTAX, Allschwill, Switzerland),mounting the cell onto its apparatus and then pumping the appropriatemedium (typically FaSSIF, FeSSIF, SGF) through the powder. Thetemperature of the apparatus is typically set to 37° C. The amount ofpowder added into the cell depends on drug load of the powder: The exactamount of powder can be calculated from results obtained from drug loadanalysis of the powders. The PKI may be added into the flow through celland a flow rate between 5 and 25 ml medium/min is pumped through thepowder. One ml samples of the medium passing through the cell iscollected at predetermined times and subsequently analyzed by HPLC (e.g.C18 column Eclipse, 4.6 mm×15 cm, 1 ml/min, detection 254 to 400 nm).Samples are typically taken after 0, 0.5, 1, 1.5, 2, 3, 4, 5, 6, 7, 8,9, 10, 15, 20, 25, 30, 35 and 40 min from the moment the medium comesout from the flow through cell. The accumulated % solubilized of theamount of active substance added into the flow through cell can becalculated and plotted against time (min). The initial slope (“initialdissolution rate”, representing 0-10 minutes) of the graph may beestimated and taken as the dissolution rate of the material in sinkcondition at 37° C. in the given dissolution medium.

Preferably, the dissolution rate is measured within the initial 0 to 10minutes of dissolution.

The increased dissolution rate is preferably measured in a solution as adissolution rate ratio of said stable, amorphous hybrid nanoparticlesand said protein kinase inhibitor in raw, crystalline form. Preferablysaid ratio is from about 1.5:1 to about 500:1, such as from about 10:1to about 30:1.

Preferably, the dissolution rate is measured in a solution withintestinal pH, such as FaSSIF or FeSSIF or in a solution with gastricpH, such as SGF.

Typically, said dissolution rate is measured by a flow through cellsystem, for instance in sink conditions. Dissolution measurement in sinkconditions of stable, amorphous hybrid nanoparticles may be measured ina method consisting of adding the wished amount of powder into a flowthrough cell system (SOTAX, Allschwill, Switzerland), mounting the cellonto its apparatus and then pumping the appropriate medium (typicallyFaSSIF, FeSSIF, SGF) through the powder. The temperature of theapparatus is typically set to 37° C. The amount of powder added into thecell depends on drug load of the powder: The exact amount of powder canbe calculated from results obtained from drug load analysis of thepowders. The PKI may be added into the flow through cell and a flow ratebetween 5 and 25 ml medium/min is pumped through the powder. One mlsamples of the medium passing through the cell is collected atpredetermined times and subsequently analyzed by HPLC (e.g. C18 columnEclipse, 4.6 mm×15 cm, 1 ml/min, detection 254 to 400 nm). Samples aretypically taken after 0, 0.5, 1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25, 30, 35 and 40 min from the moment the medium comes out from theflow through cell. The accumulated % solubilized of the amount of activesubstance added into the flow through cell, can be calculated andplotted against time (min). The initial slope (“initial dissolutionrate”, representing 0-10 minutes) of the graph can be estimated andtaken as the dissolution rate of the material in sink condition at 37°C. in the given dissolution medium.

In another embodiment of this aspect, there is provided a compositioncomprising stable, amorphous hybrid nanoparticles, comprising at leastone protein kinase inhibitor and at least one polymeric stabilizing andmatrix-forming component, which provides a solubility increase ofinhibitor in a solution, said increase measured as the area under thecurve (AUC) during about from 40 minutes to about 90 minutes, in saidsolution as compared with the AUC of inhibitor in raw, crystalline form.Typically, said increase is from about 2:1 to about 10 000:1, wherein 1represents AUC of inhibitor in raw, crystalline form. The increase maybe measured in a solution with intestinal pH, such as FaSSIF or FeSSIF,or in a solution with gastric pH, such as SGF.

The polymeric stabilizing and matrix-forming component of the presentinvention includes, but not limited to, methyl cellulose, hydroxyethylcellulose, hydroxypropyl cellulose (e.g. HPC ef, HPC lf and HPC jf),hydroxypropyl methylcellulose (e.g. Methocel E3 and E15 and Pharmacoat),hydroxypropyl methylcellulose acetate succinate (HPMC AS), hydroxypropylmethylcellulose phthalate (e.g. HPMCP HP55), polyvinylpyrrolidone (e.g.PVP 30K and PVP 90K), polyvinyl acetate phthalate (PVAP), copolyvidone(e.g. Kollidon VA 64), crospovidon (e.g. Kollidon CL), methacrylic acidand ethylacrylate copolymer (e.g. Kollicoat ME), methacrylate acid andmethyl methacrylate copolymer (e.g. Eudragit L100), polyethylene glycol(PEG), DL lactide/glycolide copolymer, poly DL-lactide, celluloseacetate phthalate (CAP), carbomer homopolymer Type A (Carbopol 971P),carbomer homopolymer Type B (Carbopol 974P), aminoalkyl methacrylatecopolymers (e.g. Eudragit RL100, RL PO or RS PO) and Poloxamers (e.g.Pluronics, Kolliphor).

Consequently, in another embodiment of this aspect, said polymericstabilizing and matrix-forming component is selected from from methylcellulose, hydroxyethyl cellulose, hydroxypropyl cellulose,hydroxypropyl methylcellulose, hydroxypropyl methylcellulose acetatesuccinate, hydroxypropyl methylcellulose phthalate,polyvinylpyrrolidone, polyvinyl acetate phthalate, copolyvidone,crospovidon, methacrylic acid and ethylacrylate copolymer, methacrylateacid and methyl methacrylate copolymer, polyethylene glycol, DLlactide/glycolide copolymer, poly DL-lactide, cellulose acetatephthalate, carbomer homopolymer Type A, carbomer homopolymer Type B,aminoalkyl methacrylate copolymers and polaxamers. Preferably, saidpolymeric stabilizing and matrix-forming component is selected fromhydroxypropyl methylcellulose phthalate, hydroxypropyl cellulose,copolyvidon, hydroxypropyl methylcellulose acetate succinate, polyvinylacetate phthalate, cellulose acetate phthalate and polyvinylpyrrolidone.

In another embodiment of this aspect, there is provided a compositioncomprising stable, amorphous hybrid nanoparticles comprising at leastone protein kinase inhibitor and at least one polymeric stabilizing andmatrix-forming component, characterized by providing an amorphous powderX-ray diffraction pattern.

In another embodiment of this aspect, there is provided a compositioncomprising stable, amorphous hybrid nanoparticles comprising at leastone protein kinase inhibitor and at least one polymeric stabilizing andmatrix-forming component, wherein the dissolution rate of said stable,amorphous hybrid nanoparticles remain stable to at least about 90%,after 6 months of storage or more, at room temperature.

In another embodiment of this aspect, said protein kinase inhibitor is atyrosine kinase inhibitor selected from the group consisting oflapatinib, pazopanib, nilotinib, erlotinib, dasatinib, gefitinib,sorafenib, crizotinib, vemurafenib and axitinib; or salts or hydrates orsolvates thereof, or combinations thereof. In some embodiments it may beadvantageous to use other PKIs. Examples of PKI include, but are notlimited to afatinib, bosutinib, cediranib, fostamatinib, imatinib,lenvatinib, lestaurtinib, motesanib, mubritinib, pegaptanib,ruxolitinib, semaxanib, sunitinib, tandunitib, tipifamib and vandetanib;or salts or hydrates or solvates thereof, or combinations thereof.

In another embodiment of this aspect, said stable, amorphous hybridnanoparticles has an average particle diameter size of less than about1000 nm, such as less than about 500 nm, preferably less than 250 nm.

In another embodiment of this aspect, said solvent is an organic solventselected from DMSO and trifluoroethanol, or a mixture of these solvents,or mixture of these solvents with other organic solvent such asDMSO/acetone, DMSO/tetrahydrofurane or trifluoroethanol/ethyl acetate.

The compositions of the present invention may also dissolve and theprotein kinase inhibitor may be systemically absorbed independently ofthe pH in the surrounding environment, and typically approximately inequal amounts, especially at both a gastric pH, such as from about pH1.2 to about pH 2.1, preferably about 1.7 and at a intestinal pH such asfrom about pH 4.5 to about pH 8, preferably at a pH of about 6. Withsystemically absorbed, is meant that the protein kinase inhibitor isreleased from the stable, amorphous hybrid nanoparticles and taken up bythe systemic blood stream. Therefore, in another embodiment of thisaspect, there is provided a composition, wherein said protein kinaseinhibitor is systemically absorbed independently of the pH. Typically,said protein kinase inhibitor is systemically absorbed withapproximately equal amounts at both a gastric pH and at an intestinalpH. Preferably, said acid pH is about pH 1.4 and preferably said neutralpH is about pH 6.5.

With approximately equal amounts is meant that the concentration ofprotein kinase inhibitor in the blood stream, after exposure isapproximately similar. This may be illustrated by a ratio, wherein theconcentration of protein kinase inhibitor in the blood stream ismeasured after administration in gastric pH conditions (A) and comparedwith the concentration of protein kinase inhibitor in the blood streamis measured after administration in intestinal pH conditions (N).Typically, the ratio A:N is from about 0.75:1 to about 1.5:1 andpreferably from about 1:1 to about 1.25:1. The concentration measurementof protein kinase inhibitor in the blood stream may be carried out as anarea under the curve (AUC) during 0-24 hours, the maximum concentration(Cmax) or as bioavailability.

Consequently, in another embodiment of this aspect, there is provided acomposition comprising stable, amorphous hybrid nanoparticles,comprising at least one protein kinase inhibitor and at least onepolymeric stabilizing and matrix-forming component, wherein theconcentration of systemically absorbed protein kinase inhibitor ingastric pH conditions compared with the concentration of systemicallyabsorbed protein kinase inhibitor in intestinal pH conditions is in aratio of from about 0.75:1 to about 1.5:1, preferably of from about 1:1to about 1.25:1. Typically said gastric pH condition represents a pH ofabout 1.4 and said intestinal pH condition represents a pH of about 6.Typically, the concentration is measured as area under the curve (AUC)during 0-24 hours of exposure of the composition or as the maximumconcentration (Cmax).

The amounts of systemically absorbed protein kinase inhibitor may bemeasured in various ways. There is provided, in Example 14 in thepresent disclosure, a method for measurement of systemically absorbedprotein kinase inhibitors at various pHs, i.e. under both acid andneutral conditions.

In another embodiment of this aspect, there is provided a compositioncomprising stable, amorphous hybrid nanoparticles, comprising at leastone protein kinase inhibitor and at least one polymeric stabilizing andmatrix-forming component, which produces a solubility increase ofinhibitor in a solution up to supersaturation, said increase measured asthe area under the curve (AUC) during about 90 minutes, in said solutionand compared with the AUC of inhibitor in crystalline form. Saidincrease may be from to about 2:1 to about 1000:1, wherein 1 representsAUC of inhibitor in crystalline form.

For understanding how the hybrid nanoparticles in the compositions ofthe invention will dissolve in vivo in the different environments of thestomach, small intestine, large intestine and colon, it is important tochoose an appropriate solution for in vitro dissolution testing. It iscritical that the in vitro test conditions mimic the in vivo environmentas closely as possible, for example pH and osmolarity. Typically, forintestinal uptake, the pH is between 6 and 7. Therefore, the solutionmay hold a pH from about pH 6 to about pH 7, such as about pH 6.5.

Therefore, in embodiments of the invention, the solution for testing hasa pH from about pH 4.5 to about pH 8, such as about pH 6.5 or such asabout pH 5. The solutions may represent Fasted Simulated StateIntestinal Fluid (FaSSIF) or Fed Simulated State Intestinal Fluid(FeSSIF).

Typically, for gastric uptake, the pH is between 1 and 2. Therefore, thesolution may hold a pH from about pH 1 to about pH 2, such as about pH1.4. Therefore, in embodiments of the invention, the solution fortesting may represent Simulated Gastric Fluid (SGF).

The choice of solution will be dependent on where in the intestinaltract and under what conditions (fasted or fed) the composition isdesired to dissolve and be taken up. Recipes and preparation of thesesolutions are obtainable from the manufacturer (Biorelevant, Croydon,U.K.). Further details are also disclosed in Jantratid, E., andDressman, J. (2009) Dissolut. Technol. 8, 21-25).

The amount of PKI in the hybrid nanoparticles in the compositions of thepresent invention may be less or more, such as wherein the amount of PKIin the hybrid nanoparticles is from about 0.01% by weight to about 99.9%by weight.

In another embodiment of this aspect, there is provided compositionscomprising stable, amorphous hybrid nanoparticles of the presentinvention, wherein the amount of PKI in the hybrid nanoparticles is fromabout 10% by weight to about 70% by weight.

In another embodiment of this aspect, there is provided providedcompositions comprising stable, amorphous hybrid nanoparticles of thepresent invention, wherein the amount of PKI in the hybrid nanoparticlesis from about 10% by weight to about 50% by weight.

In some embodiments, it may be advantagous that the amount of PKI in thestable, amorphous hybrid nanoparticles is from 5% by weight to about 50%by weight, from 10% by weight to about 40% by weight, from about 10% byweight to about 30% by weight, or from about 10% by weight to about 20%by weight.

In another embodiment of this aspect, there is provided a compositionwherein said solubilizer is present in from about 0.5% by weight toabout 95% by weight, relative to the total weight of the soliddispersion product and the solubilizer, such as at least 25% by weight,such as in at least from 25% to 65% by weight, relative to the totalweight of the solid dispersion product and the solubilizer.

Control of the characteristics of the particles may be convenient forspecific applications. Particle size, particle agglomeration, particlesporosity and the choice and ratio of the polymeric stabilizing andmatrix-forming agent could be modified in order to increase or decreasethe surface area to volume ratio of the particle or behaviour of theparticles in a gastroinstestinal fluids, leading to an increase ordecrease of the dissolution rate. Dependent on the desired dissolutioncharacteristics such particles characteristics may be adapted.Furthermore, particles with differents characteristics may be present inthe same pharmaceutical composition to provide an initial dose and aprolonged or delayed dose of active ingredient. Additionally, it may beadvantageous to provide different PKIs and/or other active ingredient(s)in different primary particles with different characteristics adapted toprovide desired dissolution rates for each active ingredient(s).

Other embodiments of the invention provide pharmaceutical compositionscomprising the stable, amorphous hybrid nanoparticles. Such compositionsmay further comprise at least one pharmaceutically acceptablesolubilizer. Said solubilizer may be present separated from the stable,amorphous hybrid nanoparticles (i.e. physically mixed with pre-preparedsolid nanoparticles) in the composition or be randomly intermixed withinthe stable, amorphous hybrid nanoparticles in the pharmaceuticalcomposition. The pharmaceutical composition may also be in a dosage formconsisting of several layers, for example laminated or multilayeredtablets, such that the hybrid nanoparticles are separated from thesolubilizer. The solubilizer may be selected from polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol copolymer,d-α-tocopherol acid polyethylene glycol 1000 succinate and ahydrogenated castor oil, such as PEG-40 hydrogenated castor oil orPEG-35 hydrogenated castor oil. Said solubilizer may also be apoloxamer.

In another embodiment of this aspect, said inhibitor is a tyrosinekinase inhibitor selected from the group consisting of lapatinib,pazopanib, nilotinib, erlotinib, dasatinib, gefitinib, sorafenibaxitinib, crizotinib and vemurafenib; or salts or hydrates or solvatesthereof, or combinations thereof.

In another embodiment of this aspect, said inhibitor is nilotinib; andsaid polymeric stabilizing and matrix-forming component is hydroxypropyl methyl cellulose phthalate or polyvinyl acetate phthalate.

In another embodiment of this aspect, said inhibitor is nilotinib; saidpolymeric stabilizing and matrix-forming component is hydroxy propylmethyl cellulose phthalate or polyvinyl acetate phthalate; and saidsolubilizer is polyvinyl caprolactam-polyvinyl acetate-polyethyleneglycol copolymer or or d-α-tocopherol acid polyethylene glycol 1000succinate.

In another embodiment of this aspect, said inhibitor is erlotinib, andsaid polymeric stabilizing and matrix-forming component is hydroxypropylmethylcellulose acetate succinate.

In another embodiment of this aspect, said inhibitor is erlotinib; saidpolymeric stabilizing and matrix-forming component is hydroxypropylmethylcellulose acetate succinate; and said solubilizer is polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol copolymer ord-α-tocopherol acid polyethylene glycol 1000 succinate.

In another embodiment of this aspect, said inhibitor is pazopanib; andsaid polymeric stabilizing and matrix-forming component ispolyvinylpyrrolidone.

In another embodiment of this aspect, said inhibitor is pazopanib; saidpolymeric stabilizing and matrix-forming component ispolyvinylpyrrolidone; and said solubilizer is polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol copolymer ord-α-tocopherol acid polyethylene glycol 1000 succinate.

In another embodiment of this aspect, said inhibitor is lapatinib; andsaid polymeric stabilizing and matrix-forming component is hydroxypropylcellulose.

In another embodiment of this aspect, said inhibitor is lapatinib; saidpolymeric stabilizing and matrix-forming component is hydroxypropylcellulose; and said solubilizer is polyvinyl caprolactam-polyvinylacetate-polyethylene glycol copolymer or d-α-tocopherol acidpolyethylene glycol 1000 succinate.

In another embodiment of this aspect, said inhibitor is gefitinib; andsaid polymeric stabilizing and matrix-forming component is hydroxypropyl methyl cellulose phthalate or polyvinyl acetate phthalate orpolyvinylpyrrolidone.

In another embodiment of this aspect, said inhibitor is gefitinib; saidpolymeric stabilizing and matrix-forming component is hydroxy propylmethyl cellulose phthalate or polyvinyl acetate phthalate orpolyvinylpyrrolidone; and said solubilizer is polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol copolymer.

In another embodiment of this aspect, said inhibitor is dasatinib; andsaid polymeric stabilizing and matrix-forming component is copolyvidone.

In another embodiment of this aspect, said inhibitor is dasatinib; saidpolymeric stabilizing and matrix-forming component is copolyvidone; andsaid solubilizer is polyvinyl caprolactam-polyvinyl acetate-polyethyleneglycol copolymer.

In another embodiment of this aspect, said inhibitor is sorafenib; andsaid polymeric stabilizing and matrix-forming component is hydroxypropyl methyl cellulose phthalate.

In another embodiment of this aspect, said inhibitor is sorafenib; saidpolymeric stabilizing and matrix-forming component is hydroxy propylmethyl cellulose phthalate; and said solubilizer is polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol copolymer.

In another embodiment of this aspect, said inhibitor is nilotinib base;and said polymeric stabilizing and matrix-forming component is hydroxypropyl methyl cellulose phthalate or polyvinyl acetate phthalate.

In another embodiment of this aspect, said inhibitor is nilotinib base;said polymeric stabilizing and matrix-forming component is hydroxypropyl methyl cellulose phthalate or polyvinyl acetate phthalate; andsaid solubilizer is polyvinyl caprolactam-polyvinyl acetate-polyethyleneglycol copolymer.

In another embodiment of this aspect, said inhibitor is axitinib; andsaid polymeric stabilizing and matrix-forming component is copolyvidoneor hydroxypropyl methylcellulose acetate succinate.

In another embodiment of this aspect, said inhibitor is axitinib; saidpolymeric stabilizing and matrix-forming component is copolyvidone orhydroxypropyl methylcellulose acetate succinate; and said solubilizer ispolyvinyl caprolactam-polyvinyl acetate-polyethylene glycol copolymer.

In another embodiment of this aspect, said inhibitor is crizotinib; andsaid polymeric stabilizing and matrix-forming component is copolyvidoneor polyvinylpyrrolidone.

In another embodiment of this aspect, said inhibitor is crizotinib; saidpolymeric stabilizing and matrix-forming component is copolyvidone orpolyvinylpyrrolidone; and said solubilizer is polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol copolymer or PEG-40hydrogenated castor oil.

In another embodiment of this aspect, said inhibitor is vemurafenib; andsaid polymeric stabilizing and matrix-forming component is copolyvidoneor cellulose acetate phthalate.

In another embodiment of this aspect, said inhibitor is vemurafenib;said polymeric stabilizing and matrix-forming component is copolyvidoneor cellulose acetate phthalate; and said solubilizer is polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol.

In another embodiment of this aspect, said protein kinase inhibitor ispartially released from the composition, at a pH of from about 1 toabout 2, preferably at about pH 1.4.

In another aspect of the invention, there is provided stable, amorphoushybrid nanoparticles, comprising at least one protein kinase inhibitorand at least one polymeric stabilizing and matrix-forming component, asdefined in the present disclosure.

In another embodiment of this aspect, there is provided a composition ofthe invention, for use in therapy.

In another embodiment of this aspect, there is provided a composition ofthe invention, for use in the treatment of proliferative disorders.Typically, said proliferative disorder is selected from tumours andcancers, including, but not limited to, neurofibromatosis, tuberoussclerosis, hemangiomas and lymphangiogenesis, cervical, anal and oralcancers, eye or ocular cancer, stomach cancer, colon cancer, bladdercancer, rectal cancer, liver cancer, pancreas cancer, lung cancer,breast cancer, cervix uteri cancer, corpus uteri cancer, ovary cancer,prostate cancer, testis cancer, renal cancer, brain cancer, cancer ofthe central nervous system, head and neck cancer, throat cancer, skinmelanoma, acute lymphocytic leukemia, acute myelogenous leukemia,Ewing's Sarcoma, Kaposi's Sarcoma, basal cell carcinoma and squamouscell carcinoma, small cell lung cancer, choriocarcinoma,rhabdomyosarcoma, angiosarcoma, hemangioendothelioma, Wilms Tumor,neuroblastoma, mouth/pharynx cancer, esophageal cancer, larynx cancer,lymphoma, multiple myeloma; cardiac hypertrophy, age-related maculardegeneration and diabetic retinopathy.

In another embodiment of this aspect, there is provided a composition ofthe invention, said composition is provided during food intake.

In another aspect of the invention, there is provided a method oftreating proliferative disorder in a patient in need thereof, comprisingadministering a therapeutically effective amount of a compositionaccording to the present invention. Said proliferative disorder istypically selected from tumours and cancers including, but not limitedto, neurofibromatosis, tuberous sclerosis, hemangiomas andlymphangiogenesis, cervical, anal and oral cancers, eye or ocularcancer, stomach cancer, colon cancer, bladder cancer, rectal cancer,liver cancer, pancreas cancer, lung cancer, breast cancer, cervix utericancer, corpus uteri cancer, ovary cancer, prostate cancer, testiscancer, renal cancer, brain cancer, cancer of the central nervoussystem, head and neck cancer, throat cancer, skin melanoma, acutelymphocytic leukemia, acute myelogenous leukemia, Ewing's Sarcoma,Kaposi's Sarcoma, basal cell carcinoma and squamous cell carcinoma,small cell lung cancer, choriocarcinoma, rhabdomyosarcoma, angiosarcoma,hemangioendothelioma, Wilms Tumor, neuroblastoma, mouth/pharynx cancer,esophageal cancer, larynx cancer, lymphoma, multiple myeloma; cardiachypertrophy, age-related macular degeneration and diabetic retinopathy.

It will be appreciated that the amount of a protein kinase inhibitor inthe stable, amorphous hybrid nanoparticles of the present inventionrequired for use in treatment will vary not only with the particularinhibitor selected but also with the route of administration, the natureof the condition for which treatment is required and the age, weight andcondition of the patient and will be ultimately at the discretion of theattendant physician. In general however a suitable dose may be in therange of from about 0.005 to about 30 mg/kg of body weight per day,preferably in the range of 0.05 to 10 mg/kg/day.

The desired dose is conveniently presented in a single dose or as adivided dose administered at appropriate intervals, for example as two,three, four or more doses per day. Dependent on the need of thetreatment and/or prevention, the desired dose may also be, for example,once every two days, once every three days, or even once a week.

The composition is conveniently administered in unit dosage form; forexample containing 0.5 to 1500 mg, conveniently 1 to 1000 mg, mostconveniently 5 to 700 mg of active ingredient per unit dosage form. Thecompositions of the invention will normally be administrated via theoral, parenteral, intravenous, intramuscular, subcutaneous or otherinjectable ways, buccal, rectal, vaginal, transdermal and/or nasal routeand/or via inhalation, in a pharmaceutically acceptable dosage form.Depending upon the disorder and patient to be treated and the route ofadministration, the compositions may be administered at varying doses.

Pharmaceutical compositions include but are not limited to thosesuitable for oral, rectal, nasal, topical (including buccal andsub-lingual), transdermal, vaginal or parenteral (includingintramuscular, subcutaneous and intravenous) administration or in a formsuitable for administration by inhalation or insufflation. Thecompositions may, where appropriate, be conveniently presented indiscrete dosage units and may be prepared by any of the methods wellknown in the art of pharmacy. Pharmaceutical compositions suitable fororal administration are conveniently presented as discrete units such ascapsules, cachets or tablets, each containing a predetermined amount ofthe active substance.

Tablets and capsules for oral administration may contain conventionalexcipients such as binding agents, fillers, lubricants, disintegrants,or wetting agents. The tablets may be coated according to methods wellknown in the art.

The compositions may be formulated for parenteral administration (e.g.by injection, for example bolus injection or continuous infusion) andmay be presented in unit dose form in ampoules, pre-filled syringes,small volume infusion or in multi-dose containers with an addedpreservative. The compositions may take such forms as suspensions,solutions, or emulsions in oily or aqueous vehicles, and may containformulation agents such as suspending, stabilizing and/or dispersingagents.

The above described compositions may be adapted to give sustainedrelease of the active inhibitor.

The following examples are provided to illustrate various embodiments ofthe present invention and shall not be considered as limiting in scope.

EXAMPLES

Below follows a number of non-limiting examples of compositionscomprising stable, amorphous hybrid nanoparticles. In the tables, thefollowing abbreviations to “compositions” apply:

“I” represents the protein kinase inhibitor (PKI);

“P” represents the polymeric stabilizing and matrix-forming component;

“S” represents the solubilizer;

“I+P” represents a physical mix of the inhibitor with the polymericstabilizing and matrix-forming component, i.e. without furtherprocessing;

“I+S” represents a physical mix of the inhibitor with the solubilizer;

“I+P+S” represents a physical mix of the inhibitor, the polymericstabilizing and matrix-forming component and the solubilizer;

“I/P” represents stable, amorphous hybrid nanoparticles with theinhibitor and the polymeric stabilizing and matrix-forming component;

“I/P+S” represents stable, amorphous hybrid nanoparticles with theinhibitor and the polymeric stabilizing and matrix-forming component anda separate solubilizer added; “I/P/S” represents stable, amorphoushybrid nanoparticles with the inhibitor, the polymeric stabilizing andmatrix-forming component and the solubilizer.

“Exp” represents the experiment number.

The stable, amorphous hybrid nanoparticles were produced with exemplaryPKIs, polymeric stabilizing and matrix-forming components (“Polymers”),solubilizers, solution concentrations, ratios, solvents, antisolvents,temperatures and pressures as set out below and in Table A.

A 3-6% w/v PKI/polymer solution in solvent, with a ratio PKI/polymer ofabout 20-70% w/w, was pumped through XSpray's RightSize nozzle at theflow rate of 1 ml/min using a high-performance liquid chromatographypump, together with a 100 g/min CO₂ (super- or subcritical) stream. Thepressure in the precipitation chamber was set to about 100-175 bar andthe temperature was set to about 10 to 50° C. Both streams contactwithin the nozzle and the hybrid nanoparticles were formed andsubsequently collected in the particle in the collecting chamber. TheCO₂ and solvent passed through the filtering system of the collectingchamber and were drained via the back pressure regulator outlet whichmaintains the pressure within the precipitation and collecting chambers.After pumping of the PKI/polymer solution and cleaning of the tubingwith the same solvent used to prepare the PKI/polymer solution, residualsolvents left within both the precipitation and collecting chambers wereremoved by flushing these chambers with pure scCO₂. After the flushingprocess, the CO₂ was slowly drained off from the collecting chamber.Once the CO₂ had been completely removed, the particles on the filteringsystem were collected for analysis.

For I/P/S type particles, a defined amount of solubilizer is added anddissolved into the PKI/polymer solution before pumping the solutionthrough the nozzle for precipitation according to the methods describedabove.

For I/P+S type particles, a defined amount of solubilizer is added tothe stable, amorphous hybrid nanoparticles in a glass vial. The glassvial is slowly rotated for mixing of the solubilizer with the hybridnanoparticles.

TABLE A Stable, amorphous hybrid nanoparticles with exemplary PKIs,polymeric stabilizing and matrix-forming components, solvents,antisolvents and conditions. Solution Ratio conc. % PKI/Polymer SolventTemperature PKI/Polymer Exp. # (w/v) % (w/w) & Antisolvent & PressureAxitinib/ 160, 162 & 5% 25% DMSO 25° C. Kollidon 581 & CO₂ & 125 BarsVA64 Crizotinib/ 153, 155, 5% 25% DMSO 25° C. PVP 30K 156 & 571 & CO₂ &125 Bars Dasatinib/ 140, 141 & 4% 35% DMSO/Acetone 15° C. Kollidon 551(1:2) & CO₂ & 125 Bars VA64 Erlotinib HCl/ 511 3.6%   35% TFE 25° C.HPMC AS & CO₂ & 150 Bars Gefitinib/ 135, 137 & 4% 35% DMSO/Acetone 40°C. HPMCP HP55 541 (1:2) & CO2 & 150 Bars Lapatinib base/ 531 5% 66%DMSO/Acetone 40° C. HPC If (1:2) & CO2 & 150 Bars Nilotinib base/ 501 5%40% TFE 15° C. HPMCP HP55 & CO₂ & 125 Bars Pazopanib HCl/ 521 3.6%   35%TFE 25° C. PVP 90K & CO₂ & 150 Bars Sorafenib 561 4% 35% DMSO/Acetone40° C. tosylate/ (1:2) & CO2 & 150 Bars HPMCP HP55 Vemurafenib/ 168, 170& 5% 25% DMSO 25° C. CAP 592 & CO₂ & 125 Bars

General Description of Dissolution Measurement Assay

The method consists of adding the wished amount of powder of stable,amorphous hybrid nanoparticles into a glass vial and then pouring in itthe appropriate medium (typically FaSSIF, FeSSIF or SGF). The medium wasprepared in accordance with the manufacturer's instructions. The amountof powder added depends on the wished “total PKI concentration”. Forsome experiments where powders with high drug loads were tested andcompared, the real amount of PKI in the stable, amorphous hybridnanoparticles was not taken in account. For other experiments, the drugload was first estimated by HPLC and the amount of powder to obtain thedrug concentration was calculated.

Typically, the powder was added in a 8 mL glass bottle and 7 mL ofsolution was added (typically FaSSIF, FeSSIF or SGF). The glass bottlewas put on a shaker (approximately 1 rotation per minute) fordissolution. Samples of 500 μl where taken after different times, andsubsequently centrifuged at approximately 15000 g for 3 minutes. Theresulting supernatant was then analyzed by HPLC (C18 column Eclipse, 4.6mm—15 cm, 1 mL/min, detection at 254-400 nM. Generally samples weretaken after 5, 30 and 90 min and eventually 150 min.

Example 1. Compositions with Stable, Amorphous Hybrid Nanoparticles withNilotinib—Wolubility at pH 6.5 and pH 5

A number of experiments were carried out, wherein nilotinib base ornilotinib HCl represented the protein kinase inhibitor. The experimentswere carried out by measuring concentration of solubilized PKI (mg/L)after 5, 30 and 90 minutes dissolution in a solution at about pH 6.5,namely FaSSIF (Fasted State Simulated Intestinal Fluid). Further,experiments were carried out in an alternative solution at about pH 5,namely FeSSIF (Fed State Simulated Intestinal Fluid). Samples of thesolution were taken at various time intervals and the amount of proteinkinase inhibitor was measured by the dissolution measurement assaydescribed above.

Representative results in FaSSIF solution are provided below in Table 1and 2, where Table 1 provides data of concentration of nilotinib HCl(mg/L) after 5, 30 and 90 minutes dissolution, whereas Table 2 providesdata of % solubilized nilotinib HCl after 30 minutes dissolution, theArea Under the Curve (AUC—mg/min/L) during 90 minutes dissolution andthe AUC increase of stable, amorphous hybrid nanoparticles, compared tonilotinib HCl in raw, crystalline form added to the solution(experiments 1-40). In Tables 3 and 4, there is provided dissolutiondata in FeSSIF solution, presented similarly as Table 1 and 2(experiments 41-55). Table 5 provides data from a comparative experimentwith similar stable, amorphous hybrid nanoparticles, carried out inFaSSIF and FeSSIF, respectively (experiments 56-57). Table 6 presentsfurther comparative data for experiments carried out in FaSSIF andFeSSIF, respectively.

TABLE 1 Nilotinib - concentration of nilotinib HCl (mg/L) after 5, 30and 90 minutes dissolution in FaSSIF solution (pH 6.5). Drug Polymericload stab. matrix. Conc Conc Conc ratio Component Solubilizer (mg/L)(mg/L) (mg/L) Exp Comp. Inhibitor (I) (%) (P) (S) 5 min 30 min 90 min 1I Nilotinib HCl 100 — — 0.1 0.2 0.1 (raw) 100 mg 2 I Nilotinib HCl 100 —— 0.2 0.2 0.2 (raw) 500 mg 3 I Nilotinib HCl 100 — — 0.2 0.3 0.2 (raw)1000 mg 4 I Nilotinib Base 100 — — 0.6 0.5 0.2 (raw) 500 mg 5 I + PNilotinib HCl 100 HPMCP HP55 — 0.2 0.5 0.5 (raw) 1000 mg 2000 mg 6 I + PNilotinib HCl 100 PVAP — 1.3 0.2 0.4 (raw) 1000 mg 2000 mg 7 I + PNilotinib HCl 100 Eudragit L100 — 0.2 0.4 0.2 (raw) 1000 mg 2000 mg 8I + P Nilotinib HCl 100 Methocel E15 — 0.1 0.1 0.1 (raw) 1000 mg 2000 mg9 I + S Nilotinib HCl 100 — Soluplus 0.4 0.3 0.4 (raw) 1000 mg 357.5 mg10 I + S Nilotinib HCl 100 — Soluplus 0.4 0.5 0.5 (raw) 1000 mg 715 mg11 I + S Nilotinib HCl 100 — Soluplus 0.4 0.5 0.6 (raw) 1000 mg 1072 mg12 I + P + S Nilotinib HCl 100 HPMCP HP55 Soluplus 0.4 0.6 1.0 (raw) 500mg 750 mg 715 mg 13 I + P + S Nilotinib HCl 100 PVAP Soluplus 0.2 0.20.3 (raw) 500 mg 750 mg 715 mg 14 I + P + S Nilotinib HCl 100 HPMCP HP55TPGS 0.5 0.9 1.1 (raw) 500 mg 750 mg 1000 mg 15 I + P + S Nilotinib HCl100 PVAP TPGS 0.2 0.4 0.5 (raw) 500 mg 750 mg 1000 mg 16 I + P + SNilotinib Base 100 HPMCP HP55 Soluplus 0.2 0.5 0.4 (raw) 500 mg 750 mg715 mg 17 I/P Nilotinib HCl 50 HPMCP HP55 — 9.5 5.6 4.5 100 mg 100 mg 18I/P Nilotinib HCl 40 HPMCP HP55 — 10.4 5.0 3.7 100 mg 150 mg 19 I/PNilotinib HCl 50 PVAP — 7.3 5.0 4.1 100 mg 100 mg 20 I/P Nilotinib HCl40 PVAP — 8.7 5.0 3.4 100 mg 150 mg 21 I/P Nilotinib HCl 50 Methocel E15— 1.4 1.5 1.8 100 mg 100 mg 22 I/P Nilotinib HCl 50 Eudragit L100 — 5.15.9 4.9 100 mg 100 mg 23 I/P Nilotinib Base 40 HPMCP HP55 — 9.7 4.7 3.8100 mg 150 mg 24 I/P + S Nilotinib HCl 50 HPMCP HP55 Soluplus 53.4 46.135.6 500 mg 500 mg 715 mg 25 I/P + S Nilotinib HCl 50 HPMCP HP55Soluplus 85.9 87.9 80.8 500 mg 500 mg 1430 mg 26 I/P + S Nilotinib HCl50 HPMCP HP55 Soluplus 117.0 127.1 116.9 500 mg 500 mg 2145 mg 27 I/P +S Nilotinib HCl 50 HPMCP HP55 TPGS 49.6 30.1 22.3 500 mg 500 mg 1000 mg28 I/P + S Nilotinib HCl 50 HPMCP HP55 TPGS 98.4 57.4 42.6 500 mg 500 mg2000 mg 29 I/P + S Nilotinib HCl 40 HPMCP HP55 Soluplus 93.5 45.2 14.1500 mg 750 mg 357.5 mg 30 I/P + S Nilotinib HCl 40 HPMCP HP55 Soluplus145.0 134.3 36.8 500 mg 750 mg 715 mg 31 I/P + S Nilotinib HCl 40 HPMCPHP55 TPGS 93.8 31.0 22.4 500 mg 750 mg 1000 mg 32 I/P + S Nilotinib HCl40 PVAP Soluplus 82.9 137.9 42.9 500 mg 750 mg 715 mg 33 I/P + SNilotinib HCl 40 PVAP TPGS 77.8 32.3 22.8 500 mg 750 mg 1000 mg 34 I/P +S Nilotinib HCl 50 Methocel E15 Soluplus 3.3 4.0 5.8 500 mg 500 mg 715mg 35 I/P + S Nilotinib HCl 50 Methocel E15 TPGS 4.8 5.4 6.7 500 mg 500mg 1000 mg 36 I/P + S Nilotinib Base 40 HPMCP HP55 Soluplus 178.1 120.433.7 500 mg 750 mg 715 mg 37 I/P/S Nilotinib HCl 25.4 HPMCP HP55Soluplus 25.9 15.8 16.3 500 mg 750 mg 715 mg 38 I/P/S Nilotinib HCl 25.4PVAP Soluplus 9.5 13.2 10.1 500 mg 750 mg 715 mg 39 I/P/S Nilotinib HCl22.2 HPMCP HP55 TPGS 16.2 13.7 3.9 500 mg 750 mg 1000 mg 40 I/P/SNilotinib HCl 22.2 PVAP TPGS 13.3 12.1 9.7 500 mg 750 mg 1000 mg

TABLE 2 Percentage solubilized nilotinib HCl after 30 minutesdissolution, the Area Under the Curve (AUC - mg/min/L) during 90 minutesdissolution and the AUC increase of compositions comprising stable,amorphous hybrid nanoparticles with nilotinib, compared to nilotinib HClin raw, crystalline form added to the FaSSIF solution (pH 6.5). DrugPolymeric load stab. matrix. AUC/ ratio Component Solubilizer %solubilized 90 min AUC Exp Comp. Inhibitor (I) (%) (P) (S) 30 min.Mg/min/L increase 1 I Nilotinib HCl 100 — — 0.20 13.0 — (raw) 100 mg 2 INilotinib HCl 100 — — 0.04 23.5 — (raw) 500 mg 3 I Nilotinib HCl 100 — —0.03 21.8 — (raw) 1000 mg 4 I Nilotinib Base 100 — — 0.50 36.5 — (raw)500 mg 5 I + P Nilotinib HCl 100 HPMCP HP55 — 0.05 39.3 2.0 (raw) 1000mg 2000 mg 6 I + P Nilotinib HCl 100 PVAP 2000 mg — 0.02 40.0 2.1 (raw)1000 mg 7 I + P Nilotinib HCl 100 Eudragit L100 — 0.04 26.0 1.3 (raw)1000 mg 2000 mg 8 I + P Nilotinib HCl 100 Methocel E15 — 0.01 8.8 0.5(raw) 1000 mg 2000 mg 9 I + S Nilotinib HCl 100 — Soluplus 0.03 30.8 1.6(raw) 1000 mg 357.5 mg 10 I + S Nilotinib HCl 100 — Soluplus 0.05 42.32.2 (raw) 1000 mg 715 mg 11 I + S Nilotinib HCl 100 — Soluplus 0.05 45.32.3 (raw) 1000 mg 1072 mg 12 I + P + S Nilotinib HCl 100 HPMCP HP55Soluplus 0.12 61.5 3.2 (raw) 500 mg 750 mg 715 mg 13 I + P + S NilotinibHCl 100 PVAP Soluplus 0.04 20.5 1.1 (raw) 500 mg 750 mg 715 mg 14 I +P + S Nilotinib HCl 100 HPMCP HP55 TPGS 0.18 78.8 4.1 (raw) 500 mg 750mg 1000 mg 15 I + P + S Nilotinib HCl 100 PVAP TPGS 0.08 35.0 1.8 (raw)500 mg 750 mg 1000 mg 16 I + P + S Nilotinib Base 100 HPMCP HP55Soluplus 0.10 36.3 1.9 (raw) 500 mg 750 mg 715 mg 17 I/P Nilotinib HCl50 HPMCP HP55 — 5.6 515.5 26.6 100 mg 100 mg 18 I/P Nilotinib HCl 40HPMCP HP55 — 5.0 479.5 24.7 100 mg 150 mg 19 I/P Nilotinib HCl 50 PVAP —5.0 445.0 22.9 100 mg 100 mg 20 I/P Nilotinib HCl 40 PVAP — 5.0 445.022.9 100 mg 150 mg 21 I/P Nilotinib HCl 50 Methocel E15 — 1.5 138.8 7.2100 mg 100 mg 22 I/P Nilotinib HCl 50 Eudragit L100 — 5.9 474.3 24.2 100mg 100 mg 23 I/P Nilotinib Base 40 HPMCP HP55 — 4.7 459.3 23.7 100 mg150 mg 24 I/P + S Nilotinib HCl 50 HPMCP HP55 Soluplus 9.2 3828.3 197.3500 mg 500 mg 715 mg 25 I/P + S Nilotinib HCl 50 HPMCP HP55 Soluplus17.6 7448.3 383.9 500 mg 500 mg 1430 mg 26 I/P + S Nilotinib HCl 50HPMCP HP55 Soluplus 25.4 10663.8 549.7 500 mg 500 mg 2145 mg 27 I/P + SNilotinib HCl 50 HPMCP HP55 TPGS 6.0 2692.3 138.8 500 mg 500 mg 1000 mg28 I/P + S Nilotinib HCl 50 HPMCP HP55 TPGS 11.5 5193.5 267.7 500 mg 500mg 2000 mg 29 I/P + S Nilotinib HCl 40 HPMCP HP55 Soluplus 9.0 3746.5193.1 500 mg 750 mg 357.5 mg 30 I/P + S Nilotinib HCl 40 HPMCP HP55Soluplus 26.9 8974.8 462.6 500 mg 750 mg 715 mg 31 I/P + S Nilotinib HCl40 HPMCP HP55 TPGS 6.2 3396.5 175.1 500 mg 750 mg 1000 mg 32 I/P + SNilotinib HCl 40 PVAP Soluplus 27.6 8391.3 432.5 500 mg 750 mg 715 mg 33I/P + S Nilotinib HCl 40 PVAP TPGS 6.5 3223.8 166.2 500 mg 750 mg 1000mg 34 I/P + S Nilotinib HCl 50 Methocel E15 Soluplus 0.8 393.5 20.3 500mg 500 mg 715 mg 35 I/P + S Nilotinib HCl 50 Methocel E15 TPGS 1.1 505.525.9 500 mg 500 mg 1000 mg 36 I/P + S Nilotinib Base 40 HPMCP HP55Soluplus 24.1 8799.5 453.6 500 mg 750 mg 715 mg 37 I/P/S Nilotinib HCl25.4 HPMCP HP55 Soluplus 3.2 1549.0 79.8 500 mg 750 mg 715 mg 38 I/P/SNilotinib HCl 25.4 PVAP Soluplus 2.6 1006.5 51.9 500 mg 750 mg 715 mg 39I/P/S Nilotinib HCl 22.2 HPMCP HP55 TPGS 2.7 942.3 48.6 500 mg 750 mg1000 mg 40 I/P/S Nilotinib HCl 22.2 PVAP TPGS 2.4 1004.8 51.8 500 mg 750mg 1000 mg

TABLE 3 Nilotinib - concentration of nilotinib HCl (mg/L) after 5, 30and 90 minutes dissolution in FeSSIF solution (pH 5). Drug Polymericload stab. matrix. Conc Conc Conc ratio Component Solubilizer (mg/L)(mg/L) (mg/L) Exp Comp. Inhibitor (I) (%) (P) (S) 5 min 30 min 90 min 41I Nilotinib HCl 100 — — 0.6 0.9 0.9 (raw) 500 mg 42 I + P + S NilotinibHCl 100 HPMCP HP55 Soluplus 0.4 0.6 1.0 (raw) 500 mg 750 mg 715 mg 43I + P + S Nilotinib HCl 100 HPMCP HP55 Soluplus 0.2 0.2 0.3 (raw) 500 mg750 mg 1000 mg 44 I + P + S Nilotinib HCl 100 PVAP Soluplus 0.5 0.9 1.1(raw) 500 mg 750 mg 715 mg 45 I + P + S Nilotinib HCl 100 PVAP TPGS 0.20.4 0.5 (raw) 500 mg 750 mg 1000 mg 46 I/P Nilotinib HCl 40 HPMCP HP55 —16.2 45.6 63.3 500 mg 750 mg 47 I/P Nilotinib HCl 40 PVAP — 3 7.7 11.2500 mg 150 mg 48 I/P + S Nilotinib HCl 40 HPMCP HP55 Soluplus 47.7 85.5109.4 500 mg 750 mg 715 mg 49 I/P + S Nilotinib HCl 40 HPMCP HP55 TPGS74.8 112.4 125.5 500 mg 750 mg 1000 mg 50 I/P + S Nilotinib HCl 40 PVAPSoluplus 12.9 21.3 27.3 500 mg 750 mg 715 mg 51 I/P + S Nilotinib HCl 40PVAP TPGS 20.5 29.8 31.8 500 mg 750 mg 1000 mg 52 I/P/S Nilotinib HCl 40HPMCP HP55 Soluplus 42.3 81.5 108.1 500 mg 750 mg 715 mg 53 I/P/SNilotinib HCl 40 HPMCP HP55 TPGS 86.3 116.3 128.8 500 mg 750 mg 1000 mg54 I/P/S Nilotinib HCl 40 PVAP Soluplus 6.3 18.8 28.2 500 mg 750 mg 715mg 55 I/P/S Nilotinib HCl 40 PVAP TPGS 20.5 29.8 31.8 500 mg 750 mg 1000mg

TABLE 4 Percentage solubilized nilotinib HCl after 30 minutesdissolution, the Area Under the Curve (AUC - mg/min/L) during 90 minutesdissolution and the AUC increase of compositions comprising stable,amorphous hybrid nanoparticles with nilotinib, compared to nilotinib HClin raw, crystalline form added to the FeSSIF solution (pH 5). DrugPolymeric load stab. matrix. AUC/ ratio Component Solubilizer %solubilized 90 min AUC Exp Comp. Inhibitor (I) (%) (P) (S) 30 min.Mg/min/L increase 41 I Nilotinib HCl 100 — — 0.18 74.3 — (raw) 500 mg 42I + P + S Nilotinib HCl 100 HPMCP HP55 Soluplus 0.12 61.5 0.8 (raw) 500mg 750 mg 715 mg 43 I + P + S Nilotinib HCl 100 HPMCP HP55 Soluplus 0.0420.5 0.3 (raw) 500 mg 750 mg 1000 mg 44 I + P + S Nilotinib HCl 100 PVAPSoluplus 0.18 78.8 1.1 (raw) 500 mg 750 mg 715 mg 45 I + P + S NilotinibHCl 100 PVAP TPGS 0.08 35.0 0.5 (raw) 500 mg 750 mg 1000 mg 46 I/PNilotinib HCl 40 HPMCP HP55 — 9.1 4080.0 54.9 500 mg 750 mg 47 I/PNilotinib HCl 40 PVAP — 7.7 708.3 9.5 500 mg 150 mg 48 I/P + S NilotinibHCl 40 HPMCP HP55 Soluplus 17.1 7631.3 102.8 500 mg 750 mg 715 mg 49I/P + S Nilotinib HCl 40 HPMCP HP55 TPGS 22.5 9664.0 130.2 500 mg 750 mg1000 mg 50 I/P + S Nilotinib HCl 40 PVAP Soluplus 4.3 1917.8 25.8 500 mg750 mg 715 mg 51 I/P + S Nilotinib HCl 40 PVAP TPGS 6.0 2528.0 34.0 500mg 750 mg 1000 mg 52 I/P/S Nilotinib HCl 40 HPMCP HP55 Soluplus 16.37341.3 98.9 500 mg 750 mg 715 mg 53 I/P/S Nilotinib HCl 40 HPMCP HP55TPGS 23.3 10101.3 136.0 500 mg 750 mg 1000 mg 54 I/P/S Nilotinib HCl 40PVAP Soluplus 3.8 1739.5 23.4 500 mg 750 mg 715 mg 55 I/P/S NilotinibHCl 40 PVAP TPGS 6.0 2528.0 34.0 500 mg 750 mg 1000 mg

TABLE 5 Nilotinib - concentration of nilotinib HCl (mg/L) after 5, 30,90 and 150 minutes dissolution in FaSSIF and FeSSIF solution,respectively. Drug Polymeric load stab. matrix. Conc Conc Conc Concratio Component Solubilizer (mg/L) (mg/L) (mg/L) (mg/L) Exp Comp.Inhibitor (I) (%) (P) (S) 5 min 30 min 90 min 150 min 56 I/P + SNilotinib HCl 40 HPMCP HP55 Soluplus 51.2 66 62.3 53.2 FaSSIFF 75 mg112.5 mg 715 mg 57 I/P + S Nilotinib HCl 40 HPMCP HP55 Soluplus 24.843.1 50.7 53 FeSSIFF 75 mg 112.5 mg 715 mg

TABLE 6 Nilotinib - concentration of nilotinib HCl (mg/L) after 5, 30,90 and 150 minutes dissolution in FaSSIF and FeSSIF solution,respectively presented as comparative data. Drug Polymeric load stab.matrix. Compare Compare Compare Compare ratio Component Solubilizer (%)(%) (%) (%) Exp Comp. Inhibitor (I) (%) (P) (S) 5 min 30 min 90 min 150min  2 & 41 I Nilotinib HCl 100 — — 300 450 225 — (raw) 1000 mg 12 & 42I + P + S Nilotinib HCl 100 HPMCP HP55 Soluplus 100 200 250 — (raw) 750mg 715 mg 500 mg 18 & 46 I/P Nilotinib HCl 40 HPMCP HP55 — 156 912 1711— 100/500 mg 150/750 mg 30 & 48 I/P + S Nilotinib HCl 40 HPMCP HP55Soluplus 33 64 301 — 500 mg 750 mg 715 mg 56 & 57 I/P + S Nilotinib HCl40 HPMCP HP55 Soluplus 48 65 81 100 (raw) 75 mg 112.5 mg 715 mg 37 & 52I/P/S Nilotinib HCl 40 HPMCP HP55 Soluplus 163 516 663 — 500 mg 750 mg715 mg

Conclusions Example 1

Experiments 17-23 show that a solubility increase is obtained withcompositions comprising stable, amorphous hybrid nanoparticles withnilotinib HCl and a polymeric stabilizing and matrix-forming component.Particular improvements are achieved with the polymeric stabilizing andmatrix-forming components hydroxypropyl methylcellulose phthalate (HPMCPHP55) and polyvinyl acetate phthalate (PVAP). These improvements are notobtained when physically mixing nilotinib HCl with a polymericstabilizing and matrix-forming component. Experiments 24-36 clearlyshows that a further solubility increase is obtained with stable,amorphous hybrid nanoparticles with nilotinib HCl and a polymericstabilizing and matrix-forming component, wherein a separate solubilizeris added. Particular improvements are achieved by the addition of aseparate solubilizer such as polyvinyl caprolactam-polyvinylacetate-polyethylene glycol copolymer (Soluplus) or d-α-tocopherol acidpolyethylene glycol 1000 succinate (TPGS). These improvements were notobtained when physically mixing nilotinib HCl, solubilizer and/orpolymeric stabilizing and matrix-forming component (I+S or I+P+S). Noparticular improvements were obtained with stable, amorphous hybridnanoparticles with nilotinib HCl, a polymeric stabilizing andmatrix-forming and a solubilizer (I/P/S).

The results carried out in FaSSIF and FeSSIF, respectively, indicatethat the stable, amorphous hybrid nanoparticles of the invention providea similar increase in solubility. One issue with PKI formulation is thefood effect. Several of the PKIs are labeled for administration infasted state despite the fact that food in most cases increases theirbioavailability. Low bioavailability might partly explain the digestiveproblems that are associated with the PKIs. The similar dissolution ratein FaSSIF and FeSSIF indicates that the stable, amorphous hybridnanoparticles of the invention (e.g. experiments 56/57) may reduce foodeffect and patient digestive problems by its solubility improvement thatallows reducing dosage. Thus stable, amorphous hybrid nanoparticles ofthe invention may be given in conjunction with food intake.

Example 2. Compositions with Stable, Amorphous Hybrid Nanoparticles withErlotinib HCl—Solubility at pH 6.5 and pH 5

A number of experiments were carried out, wherein erlotinib HClrepresented the PKI. The experiments were carried out by measuringconcentration of PKI (mg/L) after 5, 30 and 90 minutes dissolution in asolution at about pH 6.5, namely FaSSIF (Fasted State SimulatedIntestinal Fluid). Further, experiments were carried out in analternative solution at about pH 5, namely FeSSIF (Fed State SimulatedIntestinal Fluid). Samples of the solution were taken at various timeintervals and the amount of PKI was measured by the dissolutionmeasurement assay described above.

Representative results in FaSSIF solution are provided below in Table 7and 8, where Table 7 provides data of concentration of erlotinib HCl(mg/L) after 5, 30 and 90 minutes dissolution, whereas Table 8 providesdata of % solubilized erlotinib HCl after 30 minutes dissolution, theArea Under the Curve (AUC—mg/min/L) during 90 minutes dissolution andthe AUC increase of stable, amorphous hybrid nanoparticles, compared toerlotinib HCl in raw, crystalline form added to the solution(experiments 58-68). In Tables 9 and 10, there is provided dissolutiondata in FeSSIF solution, presented similarly as Table 7 and 8(experiments 69-73). In Table 11, data from a comparative experimentwith similar stable, amorphous hybrid nanoparticles, carried out inFaSSIF and FeSSIF, respectively (experiments 74-83). Table 12 presentsfurther comparative data for experiments carried out in FaSSIF andFeSSIF, respectively.

TABLE 7 Erlotinib - concentration of erlotinib HCl (mg/L) after 5, 30and 90 minutes dissolution in FaSSIF solution (pH 6.5). Drug Polymericload stab. matrix. Conc Conc Conc ratio Component Solubilizer (mg/L)(mg/L) (mg/L) Exp Comp. Inhibitor (I) (%) (P) (S) 5 min 30 min 90 min 58I Erlotinib HCl 100 — — 28.9 6.25 4.6 (raw) 1000 mg 59 I + P ErlotinibHCl 100 HPMC-AS — 23 53.2 84 (raw) 1000 mg 2000 mg 60 I + S ErlotinibHCl 100 — Soluplus 92.8 156.6 176 (raw) 1000 mg 715 mg 61 I + SErlotinib HCl 100 — TPGS 51.4 14.7 11.6 (raw) 1000 mg 1000 mg 62 I + P +S Erlotinib HCl 100 HPMC-AS Soluplus 96.7 256.6 361.8 (raw) 1000 mg 1850mg 715 mg 63 I + P + S Erlotinib HCl 100 HPMC-AS TPGS 81.3 188.1 256.6(raw) 1000 mg 1850 mg 1000 mg 64 I/P Erlotinib HCl 35 HPMC-AS — 83.479.6 44.8 1000 mg 1850 mg 65 I/P + S Erlotinib HCl 35 HPMC-AS Soluplus187.3 269.7 284 1000 mg 1850 mg 715 mg 66 I/P + S Erlotinib HCl 35HPMC-AS TPGS 155.2 210.6 225.3 1000 mg 1850 mg 1000 mg 67 I/P/SErlotinib HCl 28 HPMC-AS Soluplus 90.1 95 96.4 1000 mg 1850 mg 715 mg 68I/P/S Erlotinib HCl 26 HPMC-AS TPGS 93.7 85.4 52.8 1000 mg 1850 mg 1000mg

TABLE 8 Percentage solubilized erlotinib HCl after 30 minutesdissolution, the Area Under the Curve (AUC - mg/min/L) during 90 minutesdissolution and the AUC increase of stable, amorphous hybridnanoparticles, compared to erlotinib in raw, crystalline form added tothe FaSSIF solution (pH 6.5). Drug Polymeric load stab. matrix. AUC/ratio Component Solubilizer % solubilized 90 min AUC Exp Comp. Inhibitor(I) (%) (P) (S) 30 min. Mg/min/L increase 58 I Erlotinib HCl 100 — — 0.6837 — (raw) 1000 mg 59 I + P Erlotinib HCl 100 HPMC-AS — 5.3 5126 6.1(raw) 1000 mg 2000 mg 60 I + S Erlotinib HCl 100 — Soluplus 15.7 1332815.9 (raw) 1000 mg 715 mg 61 I + S Erlotinib HCl 100 — TPGS 1.5 1744 2.1(raw) 1000 mg 1000 mg 62 I + P + S Erlotinib HCl 100 HPMC-AS Soluplus25.7 23210 27.7 (raw) 1000 mg 1850 mg 715 mg 63 I + P + S Erlotinib HCl100 HPMC-AS TPGS 18.8 16912 20.2 (raw) 1000 mg 1850 mg 1000 mg 64 I/PErlotinib HCl 35 HPMC-AS — 8.0 5978 7.1 1000 mg 1850 mg 65 I/P + SErlotinib HCl 35 HPMC-AS Soluplus 27.0 22792 27.2 1000 mg 1850 mg 715 mg66 I/P + S Erlotinib HCl 35 HPMC-AS TPGS 21.1 18038 21.5 1000 mg 1850 mg1000 mg 67 I/P/S Erlotinib HCl 28 HPMC-AS Soluplus 9.5 8281 9.9 1000 mg1850 mg 715 mg 68 I/P/S Erlotinib HCl 26 HPMC-AS TPGS 8.5 6619 7.9 1000mg 1850 mg 1000 mg

TABLE 9 Erlotinib - concentration of erlotinib HCl (mg/L) after 5, 30and 90 minutes dissolution in FeSSIF solution (pH 5). Drug Polymericload stab. matrix. Conc Conc Conc ratio Component Solubilizer (mg/L)(mg/L) (mg/L) Exp Comp. Inhibitor (I) (%) (P) (S) 5 min 30 min 90 min 69I Erlotinib HCl 100 — — 156.8 189.9 196 (raw) 1000 mg 70 I + P + SErlotinib HCl 100 HPMC-AS Soluplus 25.5 75.1 126.2 (raw) 1000 mg 1850 mg715 mg 71 I/P Erlotinib HCl 35 HPMC-AS — 258.2 402.1 464.5 1000 mg 1850mg 72 I/P + S Erlotinib HCl 35 HPMC-AS Soluplus 260.1 422.8 498.8 1000mg 1850 mg 715 mg 73 I/P/S Erlotinib HCl 28 HPMC-AS Soluplus 293.6 395.2434.9 1000 mg 1850 mg 715 mg

TABLE 10 Percentage solubilized erlotinib HCl after 30 minutesdissolution, the Area Under the Curve (AUC - mg/min/L) during 90 minutesdissolution and the AUC increase of stable, amorphous hybridnanoparticles, compared to erlotinib in raw, crystalline form added tothe FeSSIF solution (pH 5). Drug Polymeric load stab. matrix. AUC/ ratioComponent Solubilizer % solubilized 90 min AUC Exp Comp. Inhibitor (I)(%) (P) (S) 30 min. Mg/min/L increase 69 I Erlotinib HCl 100 — — 19.016303 — (raw) 1000 mg 70 I + P + S Erlotinib HCl 100 HPMC-AS Soluplus7.5 7360 0.5 (raw) 1000 mg 1850 mg 715 mg 71 I/P Erlotinib HCl 35HPMC-AS — 40.2 34897 2.1 1000 mg 1850 mg 72 I/P + S Erlotinib HCl 35HPMC-AS Soluplus 42.3 36835 2.3 1000 mg 1850 mg 715 mg 73 I/P/SErlotinib HCl 28 HPMC-AS Soluplus 35.5 34244 2.1 1000 mg 1850 mg 715 mg

TABLE 11 Erlotinib - concentration of erlotinib HCl after 5, 30 and 90minutes dissolution in FaSSIF and FeSSIF solution, respectively. DrugPolymeric load stab. matrix. Conc Conc Conc ratio Component Solubilizer(mg/L) (mg/L) (mg/L) Exp Comp. Inhibitor (I) (%) (P) (S) 5 min 30 min 90min 74 I + P + S Erlotinib HCl 100 HPMC-AS Soluplus 134.1 369.8 533.4FaS (raw) 1000 mg 1850 mg 1430 mg SIF 75 I + P + S Erlotinib HCl 100HPMC-AS Soluplus 24.4 88.8 154.4 FeS (raw) 1000 mg 1850 mg 1430 mg SIF76 I/P + S Erlotinib HCl 35 HPMC-AS Soluplus 275.4 441.4 508 FaS 1000 mg1850 mg 1430 mg SIF 77 I/P + S Erlotinib HCl 35 HPMC-AS Soluplus 292.2476.2 546.5 FeS 1000 mg 1850 mg 1430 mg SIF 78 I/P/S Erlotinib HCl 23HPMC-AS Soluplus 90.4 108 114.8 FaS 1000 mg 1850 mg 1430 mg SIF 79 I/P/SErlotinib HCl 23 HPMC-AS Soluplus 259.3 354.8 405.5 FeS 1000 mg 1850 mg1430 mg SIF 80 I + P + S Erlotinib HCl 100 HPMC-AS Soluplus 78.6 216.4304.6 FaS (raw) 500 mg 925 mg 715 mg SIF 81 I + P + S Erlotinib HCl 100HPMC-AS Soluplus 16.2 55.8 104.7 FeS (raw) 500 mg 925 mg 715 mg SIF 82I/P + S Erlotinib HCl 35 HPMC-AS Soluplus 171.6 284.6 334.6 FaS 500 mg925 mg 715 mg SIF 83 I/P + S Erlotinib HCl 35 HPMC-AS Soluplus 168.3268.7 317.9 FeS 500 mg 925 mg 715 mg SIF

TABLE 12 Erlotinib - concentration of erlotinib HCl (mg/L) after 5, 30and 90 minutes dissolution in FaSSIF and FeSSIF solution, respectivelypresented as comparative data. Drug Polymeric load stab. matrix. CompareCompare Compare ratio Component Solubilizer (%) (%) (%) Exp Comp.Inhibitor (I) (%) (P) (S) 5 min 30 min 90 min 58 & 69 I Erlotinib HCl100 — — 543 3038 4261 (raw) 1000 mg 74 & 75 I + P + S Erlotinib HCl 100HPMC-AS Soluplus 18 24 29 (raw) 1000 mg 1850 mg 1430 mg 80 & 81 I + P +S Erlotinib HCl 100 HPMC-AS Soluplus 21 26 34 (raw) 925 mg 715 mg 500 mg64 & 71 I/P Erlotinib HCl 35 HPMC-AS — 310 505 1037 1000 mg 1850 mg 76 &77 I/P + S Erlotinib HCl 35 HPMC-AS Soluplus 106 108 108 1000 mg 1850 mg1430 mg 82 & 83 I/P + S Erlotinib HCl 35 HPMC-AS Soluplus 98 94 95 500mg 925 mg 715 mg 78 & 79 I/P/S Erlotinib HCl 23 HPMC-AS Soluplus 287 329353 1000 mg 1850 mg 1430 mg

Conclusions Example 2

The experiments show that a solubility increase is obtained withcompositions comprising stable, amorphous hybrid nanoparticles witherlotinib HCl and a polymeric stabilizing and matrix-forming component.Particular improvements are achieved with the polymeric stabilizing andmatrix-forming component hydroxypropyl methylcellulose acetate succinate(HPMC-AS). Experiments 65-66 and 72 show that a further solubilityincrease is obtained with stable, amorphous hybrid nanoparticles witherlotinib HCl and a polymeric stabilizing and matrix-forming component,wherein a separate solubilizer is added. Particular improvements areachieved by the addition of a separate solubilizer added, wherein saidsolubilizer is selected from polyvinyl caprolactam-polyvinylacetate-polyethylene glycol copolymer (Soluplus) and d-α-tocopherol acidpolyethylene glycol 1000 succinate (TPGS). This improvement was notobserved when the solubilizer was incorporated into the stable,amorphous hybrid nanoparticles.

Physical mixes of erlotinib HCl with a solubilizer and/or HPMC ASimprove also the solubility in FaSSIF (experiments 59, 60-61, 62-63) butnot in FeSSIF (experiment 69-72). One issue with PKI formulation is thefood effect. Several of the PKIs are labeled for administration infasted state despite the fact that food in most cases increases theirbioavailability. Low bioavailability might partly explain the digestiveproblems that are associated with the PKIs. The data indicates that thestable, amorphous hybrid nanoparticles may reduce food effect andpatient digestive problems by its equal solubility improvement in bothFaSSIF and FeSSIF (experiment 76/77 and 82/83) that moreover potentiallymay allow reducing of dosage. Thus, compositions comprising thesestable, amorphous hybrid nanoparticles may be given in conjunction withfood intake.

Example 3. Compositions with Stable, Amorphous Hybrid Nanoparticles withPazopanib—Solubility at pH 6.5 and pH 5

A number of experiments were carried out, wherein pazopanib representedthe PKI. The experiments were carried out by measuring concentration ofPKI (mg/L) after 5, 30 and 90 minutes dissolution in a solution at aboutpH 6.5, namely FaSSIF (Fasted State Simulated Intestinal Fluid).Further, experiments were carried out in an alternative solution atabout pH 5, namely FeSSIF (Fed State Simulated Intestinal Fluid).Samples of the solution were taken at various time intervals and theamount of PKI was measured by the dissolution measurement assaydescribed above.

Representative results in FaSSIF solution are provided below in Table 13and 14, where Table 13 provides data of concentration of pazopanib(mg/L) after 5, 30 and 90 minutes dissolution, whereas Table 14 providesdata of % solubilized pazopanib after 30 minutes dissolution, the AreaUnder the Curve (AUC—mg/min/L) during 90 minutes dissolution and the AUCincrease with stable, amorphous hybrid nanoparticles, compared topazopanib in raw, crystalline form added to the solution (experiments84-93). In Tables 15 and 16, there is provided dissolution data inFeSSIF solution, presented similarly as Table 13 and 14 (experiments94-101). In Table 17, data from a comparative experiment with similarstable, amorphous hybrid nanoparticles, carried out in FaSSIF andFeSSIF, respectively (experiments 102-109). Table 18 presents furthercomparative data for experiments carried out in FaSSIF and FeSSIF,respectively, with stable, amorphous hybrid nanoparticles.

TABLE 13 Pazopanib - concentration of pazopanib (mg/L) after 5, 30 and90 minutes dissolution in FaSSIF solution (pH 6.5). Drug Polymeric loadstab. matrix. Conc Conc Conc ratio Component Solubilizer (mg/L) (mg/L)(mg/L) Exp Comp. Inhibitor (I) (%) (P) (S) 5 min 30 min 90 min 84 IPazopanib (raw) 100 — — 46.2 24.4 15.0 1000 mg 85 I + P Pazopanib (raw)100 PVP 90K — 82.7 83.8 67.7 1000 mg 2000 mg 86 I + S Pazopanib (raw)100 — Soluplus 116.3 177.7 204.3 1000 mg 357 mg 87 I + S Pazopanib (raw)100 — Soluplus 177.6 270.8 324.2 1000 mg 715 mg 88 I + P + S Pazopanib(raw) 100 PVP 90K Soluplus 198.8 312.2 394.1 1000 mg 1857 mg 715 mg 89I + P + S Pazopanib (raw) 100 PVP 90K TPGS 182.6 196.7 49.2 1000 mg 1857mg 1000 mg 90 I/P Pazopanib 35 PVP 90K — 89.4 103.4 92.8 1000 mg 1857 mg91 I/P + S Pazopanib 35 PVP 90K Soluplus 238.9 409.4 469.3 1000 mg 1857mg 715 mg 92 I/P + S Pazopanib 35 PVP 90K TPGS 207.5 244.8 76.3 1000 mg1857 mg 1000 mg 93 I/P/S Pazopanib 28 PVP 90K Soluplus 127.2 128.3 82.01000 mg 1857 mg 715 mg

TABLE 14 Percentage solubilized pazopanib after 30 minutes dissolution,the Area Under the Curve (AUC - mg/min/L) during 90 minutes dissolutionand the AUC increase with stable, amorphous hybrid nanoparticles,compared to pazopanib in raw, crystalline form added to the FaSSIFsolution (pH 6.5). Drug Polymeric load stab. matrix. AUC/ ratioComponent Solubilizer % solubilized 90 min AUC Exp Comp. Inhibitor (I)(%) (P) (S) 30 min. Mg/min/L increase 84 I Pazopanib (raw) 100 — — 2.42180 — 1000 mg 85 I + P Pazopanib (raw) 100 PVP 90K — 8.4 6833 3.1 1000mg 2000 mg 86 I + S Pazopanib (raw) 100 — Soluplus 17.8 15426 7.1 1000mg 357 mg 87 I + S Pazopanib (raw) 100 — Soluplus 27.1 23899 11.0 1000mg 715 mg 88 I + P + S Pazopanib (raw) 100 PVP 90K Soluplus 31.2 2807412.9 1000 mg 1857 mg 715 mg 89 I + P + S Pazopanib (raw) 100 PVP 90KTPGS 19.7 12575 5.8 1000 mg 1857 mg 1000 mg 90 I/P Pazopanib 35 PVP 90K— 10.3 8520 3.9 1000 mg 1857 mg 91 I/P + S Pazopanib 35 PVP 90K Soluplus40.9 35062 16.1 1000 mg 1857 mg 715 mg 92 I/P + S Pazopanib 35 PVP 90KTPGS 24.5 15806 7.3 1000 mg 1857 mg 1000 mg 93 I/P/S Pazopanib 28 PVP90K Soluplus 12.8 9821 4.5 1000 mg 1857 mg 715 mg

TABLE 15 Pazopanib - concentration of pazopanib (mg/L) after 5, 30 and90 minutes dissolution in FeSSIF solution (pH 5). Drug Polymeric loadstab. matrix. Conc Conc Conc ratio Component Solubilizer (mg/L) (mg/L)(mg/L) Exp Comp. Inhibitor (I) (%) (P) (S) 5 min 30 min 90 min 94 IPazopanib (raw) 100 — — 231.3 321.4 239.3 1000 mg 95 I + P Pazopanib(raw) 100 PVP 90K — 234.8 309.7 269.7 1000 mg 2000 mg 96 I + S Pazopanib(raw) 100 — Soluplus 209.3 309.6 229.1 1000 mg 357 mg 97 I + P + SPazopanib (raw) 100 PVP 90K Soluplus 307.5 475.3 578.0 1000 mg 1857 mg715 mg 98 I + P + S Pazopanib (raw) 100 PVP 90K TPGS 320.9 395.1 325.61000 mg 1857 mg 1000 mg 99 I/P Pazopanib 35 PVP 90K — 348.4 362.1 335.81000 mg 1857 mg 100 I/P + S Pazopanib 35 PVP 90K Soluplus 450.0 684.4777.6 1000 mg 1857 mg 715 mg 101 I/P/S Pazopanib 28 PVP 90K Soluplus226.1 347.3 361.0 1000 mg 1857 mg 715 mg

TABLE 16 Percentage solubilized pazopanib after 30 minutes dissolution,the Area Under the Curve (AUC - mg/min/L) during 90 minutes dissolutionand the AUC increase with stable, amorphous hybrid nanoparticles,compared to pazopanib in raw, crystalline form added to the FeSSIFsolution (pH 5). Drug Polymeric load stab. matrix. AUC/ ratio ComponentSolubilizer % solubilized 90 min AUC Exp Comp. Inhibitor (I) (%) (P) (S)30 min. Mg/min/L increase 94 I Pazopanib (raw) 100 — — 32.1 24308 — 1000mg 95 I + P Pazopanib (raw) 100 PVP 90K — 31.0 24775 1.0 1000 mg 2000 mg96 I + S Pazopanib (raw) 100 — Soluplus 31.0 23171 1.0 1000 mg 357 mg 97I + P + S Pazopanib (raw) 100 PVP 90K Soluplus 47.5 42153 1.7 1000 mg1857 mg 715 mg 98 I + P + S Pazopanib (raw) 100 PVP 90K TPGS 39.5 313731.3 1000 mg 1857 mg 1000 mg 99 I/P Pazopanib 35 PVP 90K — 36.2 30689 1.31000 mg 1857 mg 100 I/P + S Pazopanib 35 PVP 90K Soluplus 68.4 59165 2.41000 mg 1857 mg 715 mg 101 I/P/S Pazopanib 28 PVP 90K Soluplus 34.728982 1.2 1000 mg 1857 mg 715 mg

TABLE 17 Pazopanib - concentration of pazopanib after 5, 30 and 90minutes dissolution in FaSSIF and FeSSIF solution, respectively. DrugPolymeric load stab. matrix. Conc Conc Conc ratio Component Solubilizer(mg/L) (mg/L) (mg/L) Exp Comp. Inhibitor (I) (%) (P) (S) 5 min 30 min 90min 102 I + P + S Pazopanib (raw) 100 PVP 90K Soluplus 76.8 113.8 139.6FaSSIF 300 mg 557 mg 428 mg 103 I + P + S Pazopanib (raw) 100 PVP 90KSoluplus 116.7 193.6 246.9 FeSSIF 300 mg 557 mg 428 mg 104 I/P + SPazopanib 35 PVP 90K Soluplus 154.7 214.7 223 FaSSIF 300 mg 557 mg 428mg 105 I/P + S Pazopanib 35 PVP 90K Soluplus 186 273.3 303.1 FeSSIF 300mg 557 mg 428 mg 106 I + P + S Pazopanib (raw) 100 PVP 90K Soluplus261.1 421.6 508.5 FaSSIF 1000 mg 1857 mg 1428 mg 107 I + P + S Pazopanib(raw) 100 PVP 90K Soluplus 275.8 495.4 588.0 FeSSIF 1000 mg 1857 mg 1428mg 108 I/P + S Pazopanib 35 PVP 90K Soluplus 508.9 705.8 758.4 FeSSIF1000 mg 1857 mg 1428 mg 109 I/P + S Pazopanib 35 PVP 90K Soluplus 469.1715.2 747.4 FeSSIF 1000 mg 1857 mg 1428 mg

TABLE 18 Pazopanib - concentration of pazopanib (mg/L) after 5, 30 and90 minutes dissolution in FaSSIF and FeSSIF solution, respectivelypresented as comparative data. Drug Polymeric load stab. matrix. CompareCompare Compare ratio Component Solubilizer (%) (%) (%) Exp Comp.Inhibitor (I) (%) (P) (S) 5 min 30 min 90 min 84 & 94 I Pazopanib 100 —— 501 1317 1595 (raw) 1000 mg 85 & 95 I + P Pazopanib 100 PVP 90K — 284370 398 (raw) 1857 mg 1000 mg 87 & 96 I + S Pazopanib 100 — Soluplus 118114 71 (raw) 715 mg 1000 mg 88 & 97 I + P + S Pazopanib 100 PVP 90KSoluplus 155 152 147 (raw) 1857 mg 715 mg 1000 mg 102 & 103 I + P + SPazopanib 100 PVP 90K Soluplus 152 170 177 (raw) 557 mg 428 mg 300 mg 90& 99 I/P Pazopanib 35 PVP 90K — 390 350 362 1000 mg 1857 mg  89 & 100I/P + S Pazopanib 35 PVP 90K Soluplus 188 167 166 1000 mg 1857 mg 715 mg104 & 105 I/P + S Pazopanib 35 PVP 90K Soluplus 120 127 136 300 mg 557mg 428 mg  93 & 101 I/P/S Pazopanib 28 PVP 90K Soluplus 178 271 440 1000mg 1857 mg 715 mg

Conclusions Example 3

The experiments show that a solubility increase is obtained withcompositions comprising stable, amorphous hybrid nanoparticles withpazopanib and a polymeric stabilizing and matrix-forming component.Particular improvements are achieved with the polymeric stabilizing andmatrix-forming component polyvinylpyrrolidone K-90 (PVP 90K).Experiments 91-92 show that a further solubility increase is obtainedwith stable, amorphous hybrid nanoparticles with pazopanib and apolymeric stabilizing and matrix-forming component, wherein a separatesolubilizer is added. Particular improvements are achieved by theaddition of a separate solubilizer added, wherein said solubilizer isselected from polyvinyl caprolactam-polyvinyl acetate-polyethyleneglycol copolymer (Soluplus) and d-α-tocopherol acid polyethylene glycol1000 succinate (TPGS). This improvement was not observed when thesolubilizer was incorporated into the stable, amorphous hybridnanoparticles of the invention.

The results carried out in FaSSIF and FeSSIF, respectively, indicatesthat the stable, amorphous hybrid nanoparticles of the invention providea similar increase in solubility. One issue with PKI formulation is thefood effect. Several of the PKIs are labeled for administration infasted state despite the fact that food in most cases increases theirbioavailability. Low bioavailability might partly explain the digestiveproblems that are associated with the PKIs. The similar dissolution ratein FaSSIF and FeSSIF indicates that the stable, amorphous hybridnanoparticles may reduce food effect and patient digestive problems byits equal solubility improvement in both FaSSIF and FeSSIF (experiments89/100 and 104/105) that moreover allows reducing dosage. Thus stable,amorphous hybrid nanoparticles of the invention may be given inconjunction with food intake.

Example 4. Compositions with Stable, Amorphous Hybrid Nanoparticles withLapatinib—Solubility at pH 6.5

A number of experiments were carried out, wherein lapatinib base orlapatinib ditosylate salt represented the PKI. The experiments werecarried out by measuring concentration of PKI (mg/L) after 5, 30 and 90minutes dissolution in a solution at about pH 6.5, namely FaSSIF (FastedState Simulated Intestinal Fluid). Samples of the solution were taken atvarious time intervals and the amount of PKI was measured by thedissolution measurement assay described above.

Representative results in FaSSIF solution are provided below in Table 19and 20, where Table 19 provides data of concentration of lapatinib(mg/L) after 5, 30 and 90 minutes dissolution, whereas Table 20 providesdata of % solubilized lapatinib after 30 minutes dissolution, the AreaUnder the Curve (AUC—mg/min/L) during 90 minutes dissolution and the AUCincrease with stable, amorphous hybrid nanoparticles of the invention,compared to non-formulated lapatinib ditosylate salt added to thesolution (experiments 110-126).

TABLE 19 Lapatinib - concentration of lapatinib (mg/L) after 5, 30 and90 minutes dissolution in FaSSIF solution (pH 6.5). Drug Polymeric loadstab. matrix. Conc Conc Conc ratio Component Solubilizer (mg/L) (mg/L)(mg/L) Exp Comp. Inhibitor (I) (%) (P) (S) 5 min 30 min 90 min 110 ILapatinib (base) 100 — — 2.9 6.0 6.5 2000 mg 111 I Lapatinib (salt) 100— — 57.7 132.2 124.2 2000 mg 112 I + S Lapatinib (salt) 100 — Soluplus67.6 142.9 140.0 2000 mg 285 mg 113 I + S Lapatinib (salt) 100 —Soluplus 144.7 283.6 204.0 2000 mg 645 mg 114 I + P Lapatinib (base) 100HPC LF — 1.9 4.9 6.1 2000 mg 4000 mg 115 I + P Lapatinib (salt) 100 HPCLF — 56.7 93.8 81.8 2000 mg 4000 mg 116 I + P + S Lapatinib (base) 100HPC LF Soluplus 5.5 22.5 52.0 660 mg 340 mg 715 mg 117 I + P + SLapatinib (salt) 100 HPC LF Soluplus 71.7 182.5 240.4 660 mg 340 mg 715mg 118 I + P + S Lapatinib (base) 100 HPC LF TPGS 11.8 40.6 82.9 660 mg340 mg 1000 mg 119 I + P + S Lapatinib (salt) 100 HPC LF TPGS 65.1 176.7175.3 660 mg 340 mg 1000 mg 120 I/P Lapatinib (base) 66 HPC EF — 162.5184.0 157.1 660 mg 340 mg 121 I/P Lapatinib (base) 66 HPC LF — 190.9193.5 48.0 660 mg 340 mg 122 I/P + S Lapatinib (base) 66 HPC EF Soluplus220.4 259.6 280.0 660 mg 340 mg 715 mg 123 I/P + S Lapatinib (base) 66HPC LF Soluplus 200.7 315.6 327.6 660 mg 340 mg 715 mg 124 I/P + SLapatinib (base) 66 HPC EF TPGS 202.2 237.5 242.5 660 mg 340 mg 500 mg125 I/P + S Lapatinib (base) 66 HPC LF TPGS 288.4 327.3 301.5 660 mg 340mg 500 mg 126 I/P/S Lapatinib (base) 66 HPC LF Soluplus 57.6 107.2 126.3660 mg 340 mg 715 mg

TABLE 20 Percentage solubilized lapatinib after 30 minutes dissolution,the Area Under the Curve (AUC - mg/min/L) during 90 minutes dissolutionand the AUC increase with stable, amorphous hybrid nanoparticles of theinvention, compared to non-formulated lapatinib ditosylate salt added tothe FaSSIF solution (pH 6.5). Drug Polymeric load stab. matrix. AUC/ratio Component Solubilizer % solubilized 90 min AUC Exp Comp. Inhibitor(I) (%) (P) (S) 30 min. Mg/min/L increase 110 I Lapatinib (base) 100 — —0.3 494 — 2000 mg 111 I Lapatinib (salt) 100 — — 6.6 10210 — 2000 mg 112I + S Lapatinib (salt) 100 — Soluplus 7.1 11287 1.1 2000 mg 285 mg 113I + S Lapatinib (salt) 100 — Soluplus 14.2 20344 2.0 2000 mg 645 mg 114I + P Lapatinib (base) 100 HPC LF — 0.2 420 0.04 2000 mg 4000 mg 115 I +P Lapatinib (salt) 100 HPC LF — 4.7 7291 0.7 2000 mg 4000 mg 116 I + P +S Lapatinib (base) 100 HPC LF Soluplus 3.4 2599 0.3 660 mg 340 mg 715 mg117 I + P + S Lapatinib (salt) 100 HPC LF Soluplus 27.7 16044 1.6 660 mg340 mg 715 mg 118 I + P + S Lapatinib (base) 100 HPC LF TPGS 6.2 43900.4 660 mg 340 mg 1000 mg 119 I + P + S Lapatinib (salt) 100 HPC LF TPGS26.8 13745 1.3 660 mg 340 mg 1000 mg 120 I/P Lapatinib (base) 66 HPC EF— 27.9 14971 1.5 660 mg 340 mg 121 I/P Lapatinib (base) 66 HPC LF — 29.312527 1.2 660 mg 340 mg 122 I/P + S Lapatinib (base) 66 HPC EF Soluplus39.3 22739 2.2 660 mg 340 mg 715 mg 123 I/P + S Lapatinib (base) 66 HPCLF Soluplus 47.8 26252 2.6 660 mg 340 mg 715 mg 124 I/P + S Lapatinib(base) 66 HPC EF TPGS 36.0 20402 2.0 660 mg 340 mg 500 mg 125 I/P + SLapatinib (base) 66 HPC LF TPGS 49.6 27281 2.7 660 mg 340 mg 500 mg 126I/P/S Lapatinib (base) 66 HPC LF Soluplus 16.2 9209 0.9 660 mg 340 mg715 mg

Conclusions Example 4

The experiments 122-125 clearly shows that a solubility increase isobtained with stable, amorphous hybrid nanoparticles of the invention,with lapatinib, in particular lapatinib base and a polymeric stabilizingand matrix-forming component, wherein a separate solubilizer is added tothe composition. Particular improvements are achieved with the polymericstabilizing and matrix-forming component hydroxypropyl cellulose EF andhydroxypropyl cellulose LF. Further, improvements are achieved by theaddition of a separate solubilizer added, wherein said solubilizer isselected from polyvinyl caprolactam-polyvinyl acetate-polyethyleneglycol copolymer (Soluplus) and d-α-tocopherol acid polyethylene glycol1000 succinate (TPGS).

Example 5. Compositions with Stable, Amorphous Hybrid Nanoparticles withNilotinib HCl—Solubility at pH 1.4

A number of experiments were carried out, wherein nilotinib HClrepresented the PKI. The experiments were carried out by measuringconcentration of PKI (mg/L) after 5, 30 and 90 minutes dissolution in asolution at about pH 1.4, namely SGF (Simulated Gastric Fluid). Samplesof the solution were taken at various time intervals and the amount ofPKI was measured by the dissolution measurement assay described above.

Representative results in SGF solution are provided below in Table 21,which provides percentage of solubilized nilotinib HCl from both aphysical mix with nilotinib HCl in raw, crystalline form and stable,amorphous hybrid nanoparticles of the invention after 5, 30 and 90minutes dissolution. Nilotinib present in the physical mix of nilotinibHCl raw with the polymeric stabilizing and matrix-forming component PVAPand the solubilizer Soluplus (Exp. 129) is dissolved completely within 5minutes in SGF while nilotinib is only partially dissolved after 90 minin SGF with stable, amorphous hybrid nanoparticles of the invention,wherein the components are comprised as stable, amorphous hybridnanoparticles, with the addition of a solubilizer (Exp. 128) or withoutthe addition of a solubilizer (Exp. 127).

TABLE 21 Nilotinib HCl - concentration of nilotinib HCl (mg/L) after 5,30 and 90 minutes dissolution in SGF solution (pH 1.4). Drug Polymericload stab. matrix. % % % ratio Component Solubilizer solubilizedsolubilized solubilized Exp Comp. Inhibitor (I) (%) (P) (S) 5 min 30 min90 min 127 I/P Nilotinib HCl 100 PVAP — 32 40 42 500 mg 750 mg 128 I/P +S Nilotinib HCl 100 PVAP Soluplus 38 48 50 500 mg 750 mg 715 mg 129 I +P + S Nilotinib HCl 100 PVAP Soluplus 100 100 100 (raw) 750 mg 715 mg1000 mg

Conclusions Example 5

The experiments 127-129 shows that a nilotinib HCl, in stable, amorphoushybrid nanoparticles of the invention (exp 127 and 128) are partiallysolubilized at pH 1.4. The stable, amorphous hybrid nanoparticles with apolymeric stabilizing and matrix-forming component such as PVAP ispartially protected from the acidic environment.

Example 6. Compositions with Stable, Amorphous Hybrid Nanoparticles withGefitinib—Solubility at pH 6.5

A number of experiments were carried out, wherein gefitinib representedthe PKI. The experiments were carried out by measuring concentration ofPKI (mg/L) after 3, 40 and 80 minutes dissolution in a solution at aboutpH 6.5, namely FaSSIF (Fasted State Simulated Intestinal Fluid). Samplesof the solution were taken at various time intervals and the amount ofPKI was measured by the dissolution measurement assay described above.

Representative results in FaSSIF solution are provided below in Table 22and 23, where Table 22 provides data of concentration of gefitinib(mg/L) after 3, 40 and 80 minutes dissolution, whereas Table 23 providesdata of % solubilized gefitinib after 40 minutes dissolution, the AreaUnder the Curve (AUC—mg/min/L) during 80 minutes dissolution and the AUCincrease with stable, amorphous hybrid nanoparticles of the invention,compared to non-formulated gefitinib added to the solution (experiments131-137).

TABLE 22 Gefitinib - concentration of gefitinib (mg/L) after 3, 40 and80 minutes dissolution in FaSSIF solution (pH 6.5). Drug Polymeric loadstab. matrix. Conc Conc Conc ratio Component Solubilizer (mg/L) (mg/L)(mg/L) Exp Comp. Inhibitor (I) (%) (P) (S) 3 min 40 min 80 min 131 IGefitinib (raw) 100 — — 121.8 153.1 148.1 1000 mg 132 I + P + SGefitinib (raw) 35 PVP30K Soluplus 63.6 158.3 191.1 1000 mg 1850 mg 715mg 133 I + P + S Gefitinib (raw) 35 HPMCP HP55 Soluplus 70.6 230.3 296.41000 mg 1850 mg 715 mg 134 I/P Gefitinib 35 PVP30K — 501.2 267.2 250.91000 mg 1850 mg 135 I/P Gefitinib 35 HPMCP HP55 — 254.1 321.4 332.1 1000mg 1850 mg 136 I/P + S Gefitinib 35 PVP30K Soluplus 561.4 430.2 410.91000 mg 1850 mg 715 mg 137 I/P + S Gefitinib 35 HPMCP HP55 Soluplus319.8 576.3 594.2 1000 mg 1850 mg 715 mg

TABLE 23 Percentage solubilized gefitinib after 40 minutes dissolution,the Area Under the Curve (AUC - mg/min/L) during 80 minutes dissolutionand the AUC increase with stable, amorphous hybrid nanoparticles of theinvention, compared to non-formulated gefitinib added to the FaSSIFsolution (pH 6.5). Drug Polymeric load stab. matrix. AUC/ ratioComponent Solubilizer % solubilized 80 min AUC Exp Comp. Inhibitor (I)(%) (P) (S) 40 min. Mg/min/L increase 131 I Gefitinib (raw) 100 — — 15.35967 — 1000 mg 132 I + P + S Gefitinib (raw) 35 PVP30K Soluplus 15.86630 1.1 1000 mg 1850 mg 715 mg 133 I + P + S Gefitinib (raw) 35 HPMCPHP55 Soluplus 23.0 9826 1.6 1000 mg 1850 mg 715 mg 134 I/P Gefitinib 35PVP30K — 26.7 10954 1.8 1000 mg 1850 mg 135 I/P Gefitinib 35 HPMCP HP55— 32.1 12794 2.1 1000 mg 1850 mg 136 I/P + S Gefitinib 35 PVP30KSoluplus 43.0 12282 2.9 1000 mg 1850 mg 715 mg 137 I/P + S Gefitinib 35HPMCP HP55 Soluplus 57.6 22774 3.8 1000 mg 1850 mg 715 mg

The experiments 131-137 show that a solubility increase is obtained withcompositions comprising stable, amorphous hybrid nanoparticles of theinvention, with gefitinib, in particular gefitinib and a polymericstabilizing and matrix-forming component, wherein a separate solubilizeris added to the composition. Particular improvements are achieved withthe polymeric stabilizing and matrix-forming componentpolyvinylpyrrolidone K-30 (PVP 30K) and hydroxy propyl methyl cellulosephthalate (HPMCP HP55). Further, improvements are achieved by theaddition of a separate solubilizer added, wherein said solubilizer ispolyvinyl caprolactam-polyvinyl acetate-polyethylene glycol copolymer(Soluplus).

Example 7. Compositions with Stable, Amorphous Hybrid Nanoparticles withDasatinib—Solubility at pH 6.5

A number of experiments were carried out, wherein dasatinib representedthe PKI. The experiments were carried out by measuring concentration ofPKI (mg/L) after 3, 40 and 80 minutes dissolution in a solution at aboutpH 6.5, namely FaSSIF (Fasted State Simulated Intestinal Fluid). Samplesof the solution were taken at various time intervals and the amount ofPKI was measured by the dissolution measurement assay described above.

Representative results in FaSSIF solution are provided below in Table 24and 25, where Table 24 provides data of concentration of dasatinib(mg/L) after 3, 40 and 80 minutes dissolution, whereas Table 25 providesdata of % solubilized dasatinib after 40 minutes dissolution, the AreaUnder the Curve (AUC—mg/min/L) during 80 minutes dissolution and the AUCincrease with stable, amorphous hybrid nanoparticles of the invention,compared to non-formulated dasatinib added to the solution (experiments138-141).

TABLE 24 Dasatinib - concentration of dasatinib (mg/L) after 3, 40 and80 minutes dissolution in FaSSIF solution (pH 6.5). Drug Polymeric loadstab. matrix. Conc Conc Conc ratio Component Solubilizer (mg/L) (mg/L)(mg/L) Exp Comp. Inhibitor (I) (%) (P) (S) 3 min 40 min 80 min 138 IDasatinib (raw) 100 — — 34.5 59.7 63.5 1000 mg 139 I + P + S Dasatinib(raw) 35 Kollidon VA64 Soluplus 24.2 64.9 82.5 1000 mg 1850 mg 715 mg140 I/P Dasatinib 35 Kollidon VA64 — 54.7 382.0 417.6 1000 mg 1850 mg141 I/P + S Dasatinib 35 Kollidon VA64 Soluplus 199.9 599.8 643.8 1000mg 1850 mg 715 mg

TABLE 25 Percentage solubilized dasatinib after 40 minutes dissolution,the Area Under the Curve (AUC - mg/min/L) during 80 minutes dissolutionand the AUC increase with stable, amorphous hybrid nanoparticles of theinvention, compared to non-formulated dasatinib added to the FaSSIFsolution (pH 6.5). Drug Polymeric load stab. matrix. AUC/ ratioComponent Solubilizer % solubilized 80 min AUC Exp Comp. Inhibitor (I)(%) (P) (S) 40 min. Mg/min/L increase 138 I Dasatinib (raw) 100 — — 6.02396 — 1000 mg 139 I + P + S Dasatinib (raw) 35 Kollidon VA64 Soluplus6.5 2750 1.1 1000 mg 1850 mg 715 mg 140 I/P Dasatinib 35 Kollidon VA64 —35.3 15252 6.4 1000 mg 1850 mg 141 I/P + S Dasatinib 35 Kollidon VA64Soluplus 58.6 24156 10.1 1000 mg 1850 mg 715 mg

Experiments 138-141 show that a solubility increase is obtained withcompositions comprising stable, amorphous hybrid nanoparticles of theinvention, with dasatinib, in particular dasatinib and a polymericstabilizing and matrix-forming component, wherein a separate solubilizeris added to the composition. Particular improvements are achieved withthe polymeric stabilizing and matrix-forming component copolyvidone(Kollidon VA64). Further, improvements are achieved by the addition of aseparate solubilizer added, wherein said solubilizer is polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol copolymer (Soluplus).

Example 8. Compositions with Stable, Amorphous Hybrid Nanoparticles withSorafenib Tosylate—Solubility at pH 6.5

A number of experiments were carried out, wherein sorafenib tosylaterepresented the PKI. The experiments were carried out by measuringconcentration of PKI (mg/L) after 3, 40 and 80 minutes dissolution in asolution at about pH 6.5, namely FaSSIF (Fasted State SimulatedIntestinal Fluid). Samples of the solution were taken at various timeintervals and the amount of PKI was measured by the dissolutionmeasurement assay described above.

Representative results in FaSSIF solution are provided below in Table 26and 27, where Table 26 provides data of concentration of sorafenib(mg/L) after 3, 40 and 80 minutes dissolution, whereas Table 27 providesdata of % solubilized sorafenib after 40 minutes dissolution, the AreaUnder the Curve (AUC—mg/min/L) during 80 minutes dissolution and the AUCincrease of compositions, compared to non-formulated sorafenib tosylateadded to the solution (experiments 142-145).

TABLE 26 Sorafenib tosylate - concentration of sorafenib (mg/L) after 3,40 and 80 minutes dissolution in FaSSIF solution (pH 6.5). DrugPolymeric load stab. matrix. Conc Conc Conc ratio Component Solubilizer(mg/L) (mg/L) (mg/L) Exp Comp. Inhibitor (I) (%) (P) (S) 3 min 40 min 80min 142 I Sorafenib 100 — — 59.1 343.5 311.5 tosylate (raw) 1000 mg 143I + P + S Sorafenib 35 HPMCP HP55 Soluplus 33.9 297.1 352.2 tosylate(raw) 1850 mg 715 mg 1000 mg 144 I/P Sorafenib 35 HPMCP HP55 — 245.3520.3 613.8 tosylate 1850 mg 1000 mg 145 I/P + S Sorafenib 35 HPMCP HP55Soluplus 335.1 1202.6 1738.1 tosylate 1850 mg 715 mg 2000 mg

TABLE 27 Percentage solubilized sorafenib after 40 minutes dissolution,the Area Under the Curve (AUC - mg/min/L) during 80 minutes dissolutionand the AUC increase with stable, amorphous hybrid nanoparticles of theinvention, compared to non-formulated sorafenib tosylate added to theFaSSIF solution (pH 6.5). Drug Polymeric load stab. matrix. AUC/ ratioComponent Solubilizer % solubilized 80 min AUC Exp Comp. Inhibitor (I)(%) (P) (S) 40 min. Mg/min/L increase 142 I Sorafenib 100 — — 34.4 12001— tosylate (raw) 1000 mg 143 I + P + S Sorafenib 35 HPMCP HP55 Soluplus33.9 11588 1.0 tosylate (raw) 1850 mg 715 mg 1000 mg 144 I/P Sorafenib35 HPMCP HP55 — 245.3 21838 1.8 tosylate 1850 mg 1000 mg 145 I/P + SSorafenib 35 HPMCP HP55 Soluplus 335.1 52948 4.4 tosylate 1850 mg 715 mg2000 mg

Experiments 138-141 show that a solubility increase is obtained withcompositions comprising stable, amorphous hybrid nanoparticles of theinvention, with dasatinib, in particular dasatinib and a polymericstabilizing and matrix-forming component, wherein a separate solubilizeris added to the composition. Particular improvements are achieved withthe polymeric stabilizing and matrix-forming component hydroxy propylmethyl cellulose phthalate (HPMCP HP55). Further, improvements areachieved by the addition of a separate solubilizer added, wherein saidsolubilizer is polyvinyl caprolactam-polyvinyl acetate-polyethyleneglycol copolymer (Soluplus).

Example 9. Compositions with Stable, Amorphous Hybrid Nanoparticles withNilotinib Base—Solubility at pH 6.5

A number of experiments were carried out, wherein nilotinib baserepresented the PKI. The experiments were carried out by measuringconcentration of PKI (mg/L) after 3, 40 and 80 minutes dissolution in asolution at about pH 6.5, namely FaSSIF (Fasted State SimulatedIntestinal Fluid). Samples of the solution were taken at various timeintervals and the amount of PKI was measured by the dissolutionmeasurement assay described above.

Representative results in FaSSIF solution are provided below in Table 28and 29, where Table 28 provides data of concentration of nilotinib base(mg/L) after 3, 40 and 80 minutes dissolution, whereas Table 29 providesdata of % solubilized nilotinib base after 40 minutes dissolution, theArea Under the Curve (AUC—mg/min/L) during 80 minutes dissolution andthe AUC increase of compositions, compared to non-formulated nilotinibbase added to the solution (experiments 146-149).

TABLE 28 Nilotinib base - concentration of nilotinib base (mg/L) after3, 40 and 80 minutes dissolution in FaSSIF solution (pH 6.5). DrugPolymeric load stab. matrix. Conc Conc Conc ratio Component Solubilizer(mg/L) (mg/L) (mg/L) Exp Comp. Inhibitor (I) (%) (P) (S) 3 min 40 min 80min 146 I/P Nilotinib base 40 HPMCP HP55 — 12.7 5.3 3.7 500 mg 750 mg147 I/P Nilotinib base 40 PVAP — 12.3 8.6 7.0 500 mg 750 mg 148 I/P + SNilotinib base 40 HPMCP HP55 Soluplus 136.8 88.8 41.2 500 mg 750 mg 715mg 149 I/P + S Nilotinib base 40 PVAP Soluplus 20.7 115.9 60.4 500 mg750 mg 715 mg

TABLE 29 Percentage solubilized nilotinib base after 40 minutesdissolution, the Area Under the Curve (AUC - mg/min/L) during 80 minutesdissolution and the AUC increase with stable, amorphous hybridnanoparticles of the invention, compared to non- formulated nilotinibbase added to the FaSSIF solution (pH 6.5). Drug Polymeric load stab.matrix. AUC/ ratio Component Solubilizer % solubilized 80 min AUC ExpComp. Inhibitor (I) (%) (P) (S) 40 min. Mg/min/L increase 146 I/PNilotinib base 40 HPMCP HP55 — 1.1 242 8.3 500 mg 750 mg 147 I/PNilotinib base 40 PVAP — 1.7 328 11.2 500 mg 750 mg 148 I/P + SNilotinib base 40 HPMCP HP55 Soluplus 17.8 3529 120.9 500 mg 750 mg 715mg 149 I/P + S Nilotinib base 40 PVAP Soluplus 23.2 3544 121.4 500 mg750 mg 715 mg

Example 10. Compositions with Stable, Amorphous Hybrid Nanoparticleswith Crizotinib—Solubility at pH 6.5

A number of experiments were carried out, wherein crizotinib representedthe PKI. The experiments were carried out by measuring concentration ofPKI (mg/L) after 3, 40 and 80 minutes dissolution in a solution at aboutpH 6.5, namely FaSSIF (Fasted State Simulated Intestinal Fluid). Samplesof the solution were taken at various time intervals and the amount ofPKI was measured by the dissolution measurement assay described above.

Representative results in FaSSIF solution are provided below in Table 30and 31, where Table 30 provides data of concentration of crizotinib(mg/L) after 3, 40 and 80 minutes dissolution, whereas Table 31 providesdata of % solubilized crizotinib after 40 minutes dissolution, the AreaUnder the Curve (AUC—mg/min/L) during 80 minutes dissolution and the AUCincrease with stable, amorphous hybrid nanoparticles of the invention,compared to non-formulated crizotinib added to the solution (experiments150-156).

TABLE 30 Crizotinib - concentration of crizotinib (mg/L) after 3, 40 and80 minutes dissolution in FaSSIF solution (pH 6.5). Drug Polymeric loadstab. matrix. Conc Conc Conc ratio Component Solubilizer (mg/L) (mg/L)(mg/L) Exp Comp. Inhibitor (I) (%) (P) (S) 3 min 40 min 80 min 150 ICrizotinib 100 — — 89.3 226.5 295.6 (raw) 1000 mg 151 I + P + SCrizotinib 25 PVP30K Soluplus 176.2 368.5 414.6 (raw) 1000 mg 3000 mg715 mg 152 I + P + S Crizotinib 25 PVP30K Cremophor 161.2 428.4 497.7(raw) 1000 mg 3000 mg RH40 715 mg 153 I/P Crizotinib 25 PVP30K — 325.9390.4 398.8 1000 mg 3000 mg 154 I/P Crizotinib 25 Kollidon VA64 — 297.5447.6 449.9 1000 mg 3000 mg 155 I/P + S Crizotinib 25 PVP30K Soluplus457.6 581.4 578.9 1000 mg 3000 mg 715 mg 156 I/P + S Crizotinib 25PVP30K Cremophor 573.9 855.1 867.2 1000 mg 3000 mg RH40 715 mg

TABLE 31 Percentage solubilized crizotinib after 40 minutes dissolution,the Area Under the Curve (AUC - mg/min/L) during 80 minutes dissolutionand the AUC increase with stable, amorphous hybrid nanoparticles of theinvention, compared to non-formulated crizotinib added to the FaSSIFsolution (pH 6.5). Drug Polymeric load stab. matrix. AUC/ ratioComponent Solubilizer % solubilized 80 min AUC Exp Comp. Inhibitor (I)(%) (P) (S) 40 min. Mg/min/L increase 150 I Crizotinib 100 — — 22.716773 (raw) 1000 mg 151 I + P + S Crizotinib 25 PVP30K Soluplus 36.827185 1.6 (raw) 1000 mg 3000 mg 715 mg 152 I + P + S Crizotinib 25PVP30K Cremophor 42.8 30423 1.8 (raw) 1000 mg 3000 mg RH40 715 mg 153I/P Crizotinib 25 PVP30K — 39.1 29958 1.8 1000 mg 3000 mg 154 I/PCrizotinib 25 Kollidon VA64 — 44.8 33611 2.0 1000 mg 3000 mg 155 I/P + SCrizotinib 25 PVP30K Soluplus 58.1 44862 2.7 1000 mg 3000 mg 715 mg 156I/P + S Crizotinib 25 PVP30K Cremophor 85.5 64338 3.8 1000 mg 3000 mgRH40 715 mg

Experiments 150-156 show that a solubility increase is obtained withcompositions comprising stable, amorphous hybrid nanoparticles of theinvention, with crizotinib, in particular crizotinib and a polymericstabilizing and matrix-forming component, wherein a separate solubilizeris added to the composition. Particular improvements are achieved withthe polymeric stabilizing and matrix-forming componentpolyvinylpyrrolidone K-30 (PVP 30K) and copolyvidone (Kollidon VA64).Further, improvements are achieved by the addition of a separatesolubilizer added, wherein said solubilizer is selected from polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol copolymer (Soluplus)and PEG-40 hydrogenated castor oil (Cremophor RH40).

Example 11. Compositions with Stable, Amorphous Hybrid Nanoparticleswith Axitinib—Solubility at pH 6.5

A number of experiments were carried out, wherein axitinib representedthe PKI. The experiments were carried out by measuring concentration ofPKI (mg/L) after 3, 40 and 80 minutes dissolution in a solution at aboutpH 6.5, namely FaSSIF (Fasted State Simulated Intestinal Fluid). Samplesof the solution were taken at various time intervals and the amount ofPKI was measured by the dissolution measurement assay described above.

Representative results in FaSSIF solution are provided below in Table 32and 33, where Table 32 provides data of concentration of axitinib (mg/L)after 3, 40 and 80 minutes dissolution, whereas Table 33 provides dataof % solubilized axitinib after 40 minutes dissolution, the Area Underthe Curve (AUC—mg/min/L) during 80 minutes dissolution and the AUCincrease with stable, amorphous hybrid nanoparticles of the invention,compared to non-formulated axitinib added to the solution (experiments157-163).

TABLE 32 Axitinib - concentration of axitinib (mg/L) after 3, 40 and 80minutes dissolution in FaSSIF solution (pH 6.5). Drug Polymeric loadstab. matrix. Conc Conc Conc ratio Component Solubilizer (mg/L) (mg/L)(mg/L) Exp Comp. Inhibitor (I) (%) (P) (S) 3 min 40 min 80 min 157 IAxitinib 100 — — 0.6 0.6 0.6 (raw) 500 mg 158 I + P + S Axitinib 25Kollidon VA64 Soluplus 0.2 0.8 4.3 (raw) 500 mg 1500 mg 715 mg 159 I +P + S Axitinib 25 HPMC AS Soluplus 0.2 3.0 3.1 (raw) 500 mg 1500 mg 715mg 160 I/P Axitinib 25 Kollidon VA64 — 71.1 25.9 9.1 500 mg 1500 mg 161I/P Axitinib 25 HPMC AS — 17.6 21.0 16.4 500 mg 1500 mg 162 I/P + SAxitinib 25 Kollidon VA64 Soluplus 77.6 223.6 266.1 500 mg 1500 mg 715mg 163 I/P + S Axitinib 25 HPMC AS Soluplus 40.3 110.3 129.9 500 mg 1500mg 715 mg

TABLE 33 Percentage solubilized axitinib after 40 minutes dissolution,the Area Under the Curve (AUC - mg/min/L) during 80 minutes dissolutionand the AUC increase with stable, amorphous hybrid nanoparticles of theinvention, compared to non-formulated axitinib added to the FaSSIFsolution (pH 6.5). Drug Polymeric load stab. matrix. AUC/ ratioComponent Solubilizer % solubilized 80 min AUC Exp Comp. Inhibitor (I)(%) (P) (S) 40 min. Mg/min/L increase 157 I Axitinib 100 — — 0.1 47(raw) 500 mg 158 I + P + S Axitinib 25 Kollidon VA64 Soluplus 0.2 1262.7 (raw) 500 mg 1500 mg 715 mg 159 I + P + S Axitinib 25 HPMC ASSoluplus 0.6 193 4.1 (raw) 500 mg 1500 mg 715 mg 160 I/P Axitinib 25Kollidon VA64 — 5.2 3255 69.0 500 mg 1500 mg 161 I/P Axitinib 25 HPMC AS— 4.2 1571 33.0 500 mg 1500 mg 162 I/P + S Axitinib 25 Kollidon VA64Soluplus 44.7 16070 341.0 500 mg 1500 mg 715 mg 163 I/P + S Axitinib 25HPMC AS Soluplus 22.1 7954 169.0 500 mg 1500 mg 715 mg

Experiments 157-163 show that a solubility increase is obtained withcompositions comprising stable, amorphous hybrid nanoparticles of theinvention, with axitinib, in particular axitinib and a polymericstabilizing and matrix-forming component, wherein a separate solubilizeris added to the composition. Particular improvements are achieved withthe polymeric stabilizing and matrix-forming component copolyvidone(Kollidon VA64) and hydroxypropyl methylcellulose acetate succinate(HPMC AS). Further, improvements are achieved by the addition of aseparate solubilizer added, wherein said solubilizer is polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol copolymer (Soluplus).

Example 12. Compositions with Stable, Amorphous Hybrid Nanoparticleswith Vemurafenib—Solubility at pH 6.5

A number of experiments were carried out, wherein vemurafenibrepresented the PKI. The experiments were carried out by measuringconcentration of PKI (mg/L) after 3, 40 and 80 minutes dissolution in asolution at about pH 6.5, namely FaSSIF (Fasted State SimulatedIntestinal Fluid). Samples of the solution were taken at various timeintervals and the amount of PKI was measured by the dissolutionmeasurement assay described above

Representative results in FaSSIF solution are provided below in Table 34and 35, where Table 34 provides data of concentration of vemurafenib(mg/L) after 3, 40 and 80 minutes dissolution, whereas Table 35 providesdata of % solubilized vemurafenib after 40 minutes dissolution, the AreaUnder the Curve (AUC—mg/min/L) during 80 minutes dissolution and the AUCincrease with stable, amorphous hybrid nanoparticles of the invention,compared to non-formulated vemurafenib added to the solution(experiments 164-170).

TABLE 34 Vemurafenib - concentration of vemurafenib (mg/L) after 3, 40and 80 minutes dissolution in FaSSIF solution (pH 6.5). Drug Polymericload stab. matrix. Conc Conc Conc ratio Component Solubilizer (mg/L)(mg/L) (mg/L) Exp Comp. Inhibitor (I) (%) (P) (S) 3 min 40 min 80 min164 I Vemurafenib 100 — — 0.3 0.3 0.4 (raw) 500 mg 165 I + P + SVemurafenib 25 Kollidon VA64 Soluplus 0.2 0.2 0.4 (raw) 500 mg 1500 mg715 mg 166 I + P + S Vemurafenib 25 CAP Soluplus 0.1 0.2 0.4 (raw) 500mg 1500 mg 715 mg 167 I/P Vemurafenib 25 Kollidon VA64 — 35.5 107.6122.9 500 mg 1500 mg 168 I/P Vemurafenib 25 CAP — 75.1 47.5 11.8 500 mg1500 mg 169 I/P + S Vemurafenib 25 Kollidon VA64 Soluplus 27.4 111.3172.3 500 mg 1500 mg 715 mg 170 I/P + S Vemurafenib 25 CAP Soluplus 55.4105.7 118.9 500 mg 1500 mg 715 mg

TABLE 35 Percentage solubilized vemurafenib after 40 minutesdissolution, the Area Under the Curve (AUC - mg/min/L) during 80 minutesdissolution and the AUC increase with stable, amorphous hybridnanoparticles of the invention, compared to non- formulated vemurafenibadded to the FaSSIF solution (pH 6.5). Drug Polymeric load stab. matrix.AUC/ ratio Component Solubilizer % solubilized 80 min AUC Exp Comp.Inhibitor (I) (%) (P) (S) 40 min. Mg/min/L increase 164 I Vemurafenib100 — — 0.1 27 (raw) 500 mg 165 I + P + S Vemurafenib 25 Kollidon VA64Soluplus 0.1 21 0.8 (raw) 500 mg 1500 mg 715 mg 166 I + P + SVemurafenib 25 CAP Soluplus 0.0 18 0.7 (raw) 500 mg 1500 mg 715 mg 167I/P Vemurafenib 25 Kollidon VA64 — 21.5 7669 288.0 500 mg 1500 mg 168I/P Vemurafenib 25 CAP — 9.5 3761 141.0 500 mg 1500 mg 169 I/P + SVemurafenib 25 Kollidon VA64 Soluplus 22.3 8564 322.0 500 mg 1500 mg 715mg 170 I/P + S Vemurafenib 25 CAP Soluplus 21.1 7899 297.0 500 mg 1500mg 715 mg

Experiments 164-170 show that a solubility increase is obtained withcompositions comprising stable, amorphous hybrid nanoparticles of theinvention, with vemurafenib, in particular vemurafenib and a polymericstabilizing and matrix-forming component, wherein a separate solubilizeris added to the composition. Particular improvements are achieved withthe polymeric stabilizing and matrix-forming component copolyvidone(Kollidon VA64) and cellulose acetate phthalate (CAP). Further,improvements are achieved by the addition of a separate solubilizeradded, wherein said solubilizer is polyvinyl caprolactam-polyvinylacetate-polyethylene glycol copolymer (Soluplus).

Example 13. Dissolution Rate Measurement in Sink Conditions ofCompositions of the Invention

Dissolution measurement in sink conditions of compositions of theinvention were measured in a method consisting of adding the wishedamount of powder into a flow through cell system (SOTAX, Allschwill,Switzerland), mounting the cell onto its apparatus and then pumping theappropriate medium (typically FaSSIF, FeSSIF, SGF) through the powder.The temperature of the apparatus was set to 37° C. The amount of powderadded into the cell depends on drug load of the powder: The exact amountof powder was calculated from results obtained from drug load analysisof the powders.

Typically, 3.5 to 7 mg PKI was added into the flow through cell and aflow rate between 8 and 16 ml medium/min (preferably about 8 mlmedium/min) was pumped through the powder. One ml samples of the mediumpassing through the cell were collected at predetermined times. Thesesamples were analyzed by HPLC (e.g. C18 column Eclipse, 4.6 mm×15 cm, 1ml/min, detection 254-400 nm). Samples were taken after 0, 0.5, 1, 1.5,2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35 and 40 min from themoment the medium comes out from the flow through cell. The accumulated% solubilized of the amount of active substance added into the flowthrough cell was calculated and plotted against time (min). The initialslope (“initial dissolution rate”) of the graph was estimated, asmeasured during 0 to 10 minutes, and taken as the dissolution rate ofthe material in sink condition at 37° C. in the given dissolutionmedium.

Each experiment comprises a comparison between the PKI in raw form withcompositions comprising stable, amorphous hybrid particles of theinvention with the inhibitor and a representative polymeric stabilizingand matrix-forming component.

Example 13.1. Dissolution Rate Measurement in Sink Conditions ofCompositions of the Invention, Comprising Nilotinib HCl

In experiments with nilotinib HCl, 4 mg was weighed in the flow throughcell (experiment 500) and compared with stable, amorphous hybridnanoparticles of the invention with nilotinib base and the polymericstabilizing and matrix-forming component HPMCP HP55 (experiment 501).The results are depicted in Table 36 below.

TABLE 36 Nilotinib HCl sink condition in FaSSIF. Experiment 500Experiment 501 Composition type I I/P Inhibitor (I) nilotinib HCl (raw)nilotinib base Polymeric stab. — HPMCP HP55 matrix. Component (P) Drugload % — 40% Accumulated % of solubilized of remaining active substanceat a given time (min) Min. % %  0 0.13 3.07  0.5 0.33 7.96  1 0.49 12.23 1.5 0.63 15.22  2 0.76 17.91  3 1.02 23.25  4 1.24 28.03  5 1.48 32.70 6 1.71 37.32  7 1.92 42.04  8 2.13 45.78  9 2.34 49.52 10 2.56 52.34 153.51 59.66 20 4.31 66.28 25 5.04 70.92 30 5.7 74.38 35 6.4 76.25 40 7.080.33 Initial Dissolution Rate EXP 500 0.27 EXP 501 6.58 Ratio 501/50024.0

Experiments 500-501 show that the initial dissolution rate of thestable, amorphous hybrid nanoparticles of the invention, with nilotinibbase, is superior to the initial dissolution rate of nilotinib HCl inraw, crystalline form.

Example 13.2. Dissolution Rate Measurement in Sink Conditions ofCompositions of the Invention Comprising Erlotinib HCl

In experiments with erlotinib HCl, 3.5 mg was weighed in the flowthrough cell (experiment 510) and compared with stable, amorphous hybridnanoparticles of the invention with erlotinib HCl and the polymericstabilizing and matrix-forming component HPMC AS (experiment 511). Theresults are depicted in Table 37 below.

TABLE 37 Erlotinib HCl sink condition in FaSSIF. Experiment 510Experiment 511 Composition type I I/P Inhibitor (I) erlotinib HCl (raw)erlotinib HCl Polymeric stab. — HPMC AS matrix. Component (P) Drug load% — 35% Accumulated % of solubilized of remaining active substance at agiven time (min) Min. % %  0 0.26 2.3  0.5 0.49 3.9  1 0.63 5.4  1.50.71 6.4  2 0.77 7.2  3 0.85 8.5  4 0.91 9.5  5 0.96 10.3  6 1.01 11.1 7 1.06 11.8  8 1.10 12.5  9 1.13 13.1 10 1.17 13.8 20 1.58 19.3 30 1.9322.0 40 2.24 24.6 Initial Dissolution Rate EXP 510 0.303 EXP 511 2.754Ratio 511/510 9.1

Experiments 510-511 show that the initial dissolution rate of thecompositions comprising stable, amorphous hybrid nanoparticles of theinvention, with erlotinib HCl and the polymeric stabilizing andmatrix-forming component HPMC AS, is superior to the initial dissolutionrate of erlotinib HCl in raw, crystalline form.

Example 13.3. Dissolution Rate Measurement in Sink Conditions ofCompositions of the Invention Comprising Pazopanib HCl

In experiments with pazopanib HCl, 3.5 mg was weighed in the flowthrough cell (experiment 520) and compared with stable, amorphous hybridnanoparticles of the invention with pazopanib HCl and the polymericstabilizing and matrix-forming component PVP90K (experiment 521). Theresults are depicted in Table 38 below.

TABLE 38 Pazopanib HCl sink condition in FaSSIF. Experiment 520Experiment 521 Composition type I I/P Inhibitor (I) pazopanib HCl (raw)pazopanib HCl Polymeric stab. — PVP90K matrix. Component (P) Drug load %— 35% Accumulated % of solubilized of remaining active substance at agiven time (min) Min. % %  0 1.6 1.9  0.5 4.7 4.6  1 7.7 6.8  1.5 9.68.8  2 11.2 10.6  3 13.4 15.2  4 14.7 19.7  5 15.4 22.7  6 16.0 26.2  716.4 30.1  8 16.9 33.8  9 17.2 38.2 10 17.6 41.7 20 19.2 73.2 30 20.591.3 40 21.6 97.1 Initial Dissolution Rate Dissolution Rate (5-10 min)EXP 520 4.8 0.428 EXP 521 4.33 3.85 Ratio 521/520 0.9 9.0

Experiments 520-521 show that the initial dissolution rate of thecompositions comprising stable, amorphous hybrid nanoparticles of theinvention, with pazopanib HCl and the polymeric stabilizing andmatrix-forming component PVP90K, is superior to the initial dissolutionrate of pazopanib HCl in raw, crystalline form.

Example 13.4. Dissolution Rate Measurement in Sink Conditions ofCompositions of the Invention Comprising Lapatinib Ditosylate

In experiments with lapatinib ditosylate, 4 mg was weighed in the flowthrough cell (experiment 530) and compared with stable, amorphous hybridnanoparticles of the invention with lapatinib base and the polymericstabilizing and matrix-forming component HPC lf (experiment 531). Theresults are depicted in Table 39 below.

TABLE 39 Lapatinib ditosylate sink condition in FaSSIF. Experiment 530Experiment 531 Composition type I I/P Inhibitor (I) Lapatinib ditosylate(raw) Lapatinib base Polymeric stab. — HPC If matrix. Component (P) Drugload % — 66% Accumulated % of solubilized of remaining active substanceat a given time (min) Min. % %  0 0.032 0.442  0.5 0.088 1.736  1 0.1413.053  1.5 0.190 4.448  2 0.238 5.771  3 0.332 7.504  4 0.422 8.783  50.505 9.736  6 0.582 10.573  7 0.655 11.209  8 0.725 11.732  9 0.79012.179 10 0.851 12.576 20 1.272 14.128 30 1.607 15.168 40 1.944 15.802Initial Dissolution Rate EXP 530 0.103 EXP 531 2.674 Ratio 531/530 25.9

Experiments 530-531 show that the initial dissolution rate of thecompositions comprising stable, amorphous hybrid nanoparticles of theinvention, with lapatinib base and the polymeric stabilizing andmatrix-forming component HPC lf, is superior to the initial dissolutionrate of lapatinib ditosylate in raw, crystalline form.

Example 13.5. Dissolution Rate Measurement in Sink Conditions ofCompositions of the Invention Comprising Gefitinib

In experiments with gefitinib, 3.5 mg was weighed in the flow throughcell (experiment 540) and compared with stable, amorphous hybridnanoparticles of the invention with gefitinib and the polymericstabilizing and matrix-forming component HPMCP HP55 (experiment 541).The results are depicted in Table 40 below.

TABLE 40 Gefitinib sink condition in FaSSIF. Experiment 540 Experiment541 Composition type I I/P Inhibitor (I) Gefitinib (raw) GefitinibPolymeric stab. — HPMCP HP55 matrix. Component (P) Drug load % — 35%Accumulated % of solubilized of remaining active substance at a giventime (min) Min. % %  0 0.1 1.8  0.5 0.9 6.7  1 1.9 11.3  1.5 3.2 15.4  24.5 19.0  3 7.0 23.6  4 9.5 27.4  5 11.9 30.5  6 14.3 33.5  7 16.6 36.0 8 18.8 37.8  9 20.7 39.9 10 22.7 42.6 20 29.9 50.6 30 34.1 56.7 40 36.761.8 Initial Dissolution Rate EXP 540 2.2 EXP 541 8.6 Ratio 541/540 3.9

Experiments 540-541 show that the initial dissolution rate of thecompositions comprising stable, amorphous hybrid nanoparticles of theinvention, with gefitinib and the polymeric stabilizing andmatrix-forming component HPMCP HP55, is superior to the initialdissolution rate of the gefinib in raw, crystalline form.

Example 13.6. Dissolution Rate Measurement in Sink Conditions ofCompositions of the Invention Comprising Dasatinib

In experiments with dasatinib, 3.5 mg was weighed in the flow throughcell (experiment 550) and compared with stable, amorphous hybridnanoparticles of the invention with dasatinib and the polymericstabilizing and matrix-forming component copolyvidon—Kollidon VA64(experiment 551). The results are depicted in Table 41 below.

TABLE 41 Dasatinib sink condition in FaSSIF. Experiment 550 Experiment551 Composition type I I/P Inhibitor (I) Dasatinib (raw) DasatinibPolymeric stab. — Kollidon VA64 matrix. Component (P) Drug load % — 35%Accumulated % of solubilized of remaining active substance at a giventime (min) Min. % %  0 0.3 0.4  0.5 0.7 1.0  1 1.2 1.7  1.5 1.6 2.3  22.0 2.9  3 2.8 4.2  4 3.7 5.5  5 4.4 6.8  6 5.2 8.2  7 6.0 9.5  8 6.810.8  9 7.6 12.1 10 8.3 13.4 20 15.9 25.9 30 22.1 40.9 40 26.4 54.9Initial Dissolution Rate EXP 550 0.8 EXP 551 1.3 Ratio 551/550 1.6

Experiments 550-551 show that the initial dissolution rate of thecompositions comprising stable, amorphous hybrid of the invention, withdasatinib and the polymeric stabilizing and matrix-forming componentcopolyvidon (Kollidon VA64), is superior to the initial dissolution rateof the dasatinib raw, crystalline form.

Example 13.7. Dissolution Rate Measurement in Sink Conditions ofCompositions the Invention Comprising Sorafenib Tosylate

In experiments with sorafenib tosylate, 3.5 mg was weighed in the flowthrough cell (experiment 560) and compared with stable, amorphous hybridnanoparticles of the invention with sorafenib tosylate and the polymericstabilizing and matrix-forming component HPMCP HP55 (experiment 561).The results are depicted in Table 42 below.

TABLE 42 Sorafenib tosylate sink condition in FaSSIF. Experiment 560Experiment 561 Composition type I I/P Inhibitor (I) Sorafenib tosylate(raw) Sorafenib tosylate Polymeric stab. — HPMCP HP55 matrix. Component(P) Drug load % — 35% Accumulated % of solubilized of remaining activesubstance at a given time (min) Min. % %  0 0.2 0.8  0.5 0.4 1.7  1 0.72.4  1.5 1.0 3.1  2 1.3 3.7  3 1.8 4.8  4 2.2 5.8  5 2.6 6.9  6 3.0 8.1 7 3.4 9.7  8 3.8 11.3  9 4.2 13.3 10 4.6 15.6 20 8.8 32.7 30 12.6 61.540 16.4 96.1 Initial Dissolution Rate EXP 560 0.47 EXP 561 1.17 Ratio561/560 2.5

Experiments 560-561 show that the initial dissolution rate of thecompositions comprising stable, amorphous hybrid nanoparticles of theinvention, with sorafenib tosylate and the polymeric stabilizing andmatrix-forming component HPMCP HP55, is superior to the initialdissolution rate of sorafenib tosylate in raw, crystalline form.

Example 13.8. Dissolution Rate Measurement in Sink Conditions ofCompositions of the Invention Comprising Crizotinib

In experiments with crizotinib, 3.5 mg was weighed in the flow throughcell (experiment 570) and compared with stable, amorphous hybridnanoparticles of the invention with crizotinib and the polymericstabilizing and matrix-forming component PVP 30K (experiment 571). Theresults are depicted in Table 43 below.

TABLE 43 Crizotinib sink condition in FaSSIF. Experiment 570 Experiment571 Composition type I I/P Inhibitor (I) Crizotinib (raw) CrizotinibPolymeric stab. — PVP 30K matrix. Component (P) Drug load % — 25%Accumulated % of solubilized of remaining active substance at a giventime (min) Min. % %  0 2.0 8.8  0.5 5.7 30.3  1 8.9 47.9  1.5 11.9 58.3 2 14.6 67.5  4 23.1 81.7  6 30.1 83.8  8 36.0 84.2 10 41.0 84.4 20 58.985.1 30 73.1 85.3 40 86.3 85.5 Initial Dissolution Rate EXP 570 6.6 EXP571 33.3 Ratio 571/570 5.0

Experiments 570-571 show that the initial dissolution rate of thecompositions comprising stable, amorphous hybrid nanoparticles of theinvention, with crizotinib and the polymeric stabilizing andmatrix-forming component PVP 30K, is superior to the initial dissolutionrate of crizotinib in raw, crystalline form.

Example 13.9. Dissolution Rate Measurement in Sink Conditions ofCompositions of the Invention Comprising Axitinib

In experiments with axitinib, 3.5 mg was weighed in the flow throughcell (experiment 580) and compared with stable, amorphous hybridnanoparticles of the invention with axitinib and the polymericstabilizing and matrix-forming component Kollidon VA64 (experiment 581)or HPMC AS (experiment 582). The results are depicted in Table 44 below.

TABLE 44 Axitinib sink condition in FaSSIF. Experiment Experiment 580Experiment 581 582 Composition type I I/P I/P Inhibitor (I) Axitinib(raw) Axitinib Axitinib Polymeric stab. — Kollidon VA64 HPMC AS matrix.Component (P) Drug load % — 25% 25% Accumulated % of solubilized ofremaining active substance at a given time (min) Min. % % %  0 0.03 0.750.22  0.5 0.06 1.60 0.59  1 0.08 2.33 1.04  1.5 0.11 2.97 1.50  2 0.133.56 1.92  4 0.23 6.03 3.25  6 0.31 7.76 4.39  8 0.40 9.74 5.34 10 0.4911.81 6.17 20 0.97 22.04 9.03 30 1.46 27.42 11.43 40 1.96 30.53 13.52Initial Dissolution Rate EXP 580 0.051 EXP 581 & 582 1.396 0.865 Ratio581/580 & 582/580 27.5 17.1

Experiments 580-582 show that the initial dissolution rate of thecompositions comprising stable, amorphous hybrid nanoparticles of theinvention, with axitinib and the polymeric stabilizing andmatrix-forming component Kollidon VA64 or HPMC AS, is superior to theinitial dissolution rate of axitinib in raw, crystalline form.

Example 13.10. Dissolution Rate Measurement in Sink Conditions ofCompositions of the Invention Comprising Vemurafenib

In experiments with vemurafenib, 3.5 mg was weighed in the flow throughcell (experiment 590) and compared with stable, amorphous hybridnanoparticles of the invention with vemurafenib and the polymericstabilizing and matrix-forming component Kollidon VA64 (experiment 591)or CAP (experiment 592). The results are depicted in Table 45 below.

TABLE 45 Vemurafenib sink condition in FaSSIF. Experiment Experiment 590Experiment 591 592 Composition type I I/P I/P Inhibitor (I) VemurafenibVemurafenib Vemurafenib (raw) Polymeric stab. — Kollidon VA64 CAPmatrix. Component (P) Drug load % — 25% 25% Accumulated % of solubilizedof remaining active substance at a given time (min) Min. % % %  0 0.00.1 0.4  0.5 0.0 0.2 1.1  1 0.0 0.3 1.8  1.5 0.0 0.4 2.4  2 0.0 0.5 3.1 4 0.0 1.2 6.3  6 0.0 1.9 9.4  8 0.0 2.4 11.0 10 0.0 2.9 12.1 20 0.0 4.714.9 30 0.1 5.9 16.8 40 0.1 7.1 18.3 Initial Dissolution Rate EXP 5900.002 EXP 591 & 592 0.209 1.346 Ratio 591/590 & 592/590 104 673

Experiments 590-592 clearly shows that the initial dissolution rate ofthe compositions comprising stable, amorphous hybrid nanoparticles ofthe invention, with vemurafenib and the polymeric stabilizing andmatrix-forming component Kollidon VA64 or CAP, is superior to theinitial dissolution rate of vemurafenib in raw, crystalline form.

Example 14. In Vivo Measurement of Plasma Levels After OralAdministration of Compositions of the Invention

Groups of four beagle dogs received single oral doses (5 mg/kg) ofcapsule compositions comprising stable, amorphous hybrid nanoparticlesof the invention with nilotinib base and either of the polymericstabilizing and matrix-forming components PVAP or HPMCP HP55, optionallywith addition of the solubilizer polyvinyl caprolactam-polyvinylacetate-polyethylene glycol copolymer, and compared with a marketedformulation comprising nilotinib HCl. The stable, amorphous hybridnanoparticles tested are as set out in experiments 146-149, in Example9. The stomach contents of the dogs were either neutralized with asodium bicarbonate solution 5 min prior to capsule dosing or acidifiedwith an HCl—KCl buffer 10 min prior to dose. One group of dogs alsoreceived a single iv dose (1 mg/kg) of nilotinib. Plasma levels ofnilotinib were determined with a selective LC-MS/MS method. There wereno side-effects observed in any animal studied.

Results and Conclusions

Mean±SEM plasma concentration-time profiles of nilotinib base are shownin FIGS. 22-25, and pharmacokinetic parameters and results are displayedin Tables 46A and 46B.

Outliers were calculated and excluded based on if one value is asignificant outlier from the rest at 95% confidence intervals (alpha=5%)according to Grubb's test. The critical Z value for the Grubb's test atthe 95% confidence interval with n=4 is 1.48. Z=(Mean−Questionablevalue)/SD

TABLE 46A Pharmacokinetic data following single oral administration ofdifferent nilotinib compositions of the invention, in dogs. I/P + S I/PNilotinib I/P Marketed Marketed Nilotinib base/PVAP + Nilotinib base/nilotinib nilotinib base/PVAP Soluplus PVAP formulation formulation Exp147 Exp 149 Exp 147 Acidic Neutral Acidic Acidic Neutral Stomach StomachStomach Stomach Stomach Cmax, 86 ± 52 73 ± 26 240 ± 87  360 ± 89  490 ±350 ng/mL Tmax, hr 7.6 ± 11  1.3 ± 0.3 1.3 ± 0.3 1.3 ± 0.3 1.4 ± 0.5 T½,hr 9.9; 10.7 1.9 ± 0.3 4.3 ± 3.0 3.3 ± 2.0 3.4 ± 1.2 AUC 0-24 h, 400 ±140 220 ± 90  650 ± 240 1260 ± 70  1820 ± 1200 ng * hr/mL F (%) 7.9 ±2.9 4.4 ± 1.8 13 ± 5 25 ± 1  36 ± 24

Values are given as Mean±SD, except for T ½ of the Marketed nilotinibformulation given too acid stomach where only two values were obtained.

Intravenous (IV) data were obtained by constant rate IV infusion of 1mg/kg, of a solution of Nilotinib at 0.2 mg/mL, in a 10% HPßCD, pHadjustment to pH 3.3 to 3.5. Co: 511±46 ng/mL; T½: 3.3±1.8 hr; AUCO-24hr: 1000±300 ng*hr/mL

TABLE 46B Pharmacokinetic data following single oral administration ofdifferent nilotinib formulations of the invention, in dogs. I/P I/P I/PNilotinib base/ I/P Nilotinib base/ Nilotinib base/ HPMCP HP55 +Nilotinib base/ HPMCP HP55 + HPMCP HP55 Soluplus HPMCP HP55 Soluplus Exp146 Exp 148 Exp 146 Exp 148 Acidic Acidic Neutral Stomach NeutralStomach Stomach Stomach Cmax, 210 ± 97  560 ± 220 380 ± 90  270 ± 130ng/mL Tmax, hr 1.1 ± 0.5  1.3 ± 0.29 1.2 ± 0.3 1.0 ± 0.0 T½, hr 1.9 ±0.2 3.0 ± 1.4 3.3 ± 1.3 3.8 ± 0.8 AUC 0-24 h, 730 ± 390 1600 ± 580  1230± 110  910 ± 630 ng * hr/mL F (%) 15 ± 8  32 ± 12 24 ± 2  18 ± 13

Values are given as Mean±SD

Intravenous (IV) data were obtained by constant rate IV infusion of 1mg/kg, of a solution of Nilotinib at 0.2 mg/mL, in a 10% HPßCD, pHadjustment to pH 3.3 to 3.5. Co: 511±46 ng/mL; T½: 3.3±1.8 hr; AUCO-24hr: 1000±300 ng*hr/mL.

The marketed nilotinib formulation administrated to an acidified stomachshowed plasma levels about 2 times higher than those after the sameformulation administered to a neutralized stomach. Both formulationscomprising stable, amorphous hybrid nanoparticles of the invention withnilotinib base with PVAP and HPMCP HP55 as polymeric stabilizing andmatrix-forming components showed significant improvements in plasmaexposure, with plasma levels about 2-fold higher than those of themarketed formulation given to an acidified stomach. In addition,combining stable, amorphous hybrid nanoparticles produced by the methodsof the invention could give a plasma exposure that is be more or lessindependent of stomach pH.

Further improvements in oral availability were observed whenformulations with stable, amorphous hybrid nanoparticles of theinvention were combined with the solubilizer polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol copolymer. Thus,compositions of the invention with nilotinib base with PVAP and HPMCPHP55 as polymeric stabilizing and matrix-forming components, where thesolubilizer polyvinyl caprolactam-polyvinyl acetate-polyethylene glycolcopolymer was added and administered to an acidified stomach resulted inplasma levels 2.3- to 3.1-fold higher than those of the marketedformulation. In this study, high oral bioavailability was achieved withstable, amorphous hybrid nanoparticles of the invention with nilotinibbase with HPMCP HP55 as polymeric stabilizing and matrix-formingcomponents, where the solubilizer polyvinyl caprolactam-polyvinylacetate-polyethylene glycol copolymer was added (I/P+S) andadministrated to neutralized stomach contents. In this case the exposureincreased about 7-fold over that of the marketed oral formulationadministered under the same neutralized conditions. Highestbioavalability, 36±24%, in this study was achieved when stable,amorphous hybrid nanoparticles of the invention with nilotinib base withPVAP as polymeric stabilizing and matrix-forming component wasadministered to a neutralized stomach. However, this study leg was alsoaccompanied with the highest standard deviation in the study.

There was an improvement in the in vivo performance of the compositionsof the present invention with of nilotinib stable, amorphous hybridnanoparticles, that are based on improving absorption andbioavailability by optimization of the solid state properties of thedosage form. The results of the in vivo study in dogs may predictsimilar absorption properties of stable, amorphous hybrid nanoparticlesof the invention, in patients, as there appears to be a closecorrelation in dog-human gastrointestinal drug absorption processes(Persson, E. M. et al. Pharm. Res. 2005, 22, 2141-2151). The stable,amorphous hybrid nanoparticles of the invention, with advantageousabsorption properties, also predict that the oral doses used in clinicalpractice today may be lowered. Furthermore, the stable, amorphous hybridnanoparticles of the invention may cause less pH-dependency in theabsorption and bioavailability of PKIs.

Example 15. Measurement of Degree/Level of Stability of Compositionswith Hybrid Nanoparticles of the Invention

In stability tests of compositions comprising hybrid nanoparticles ofthe invention, it was shown that particles were stable over at least 11months at room temperature (18-25° C.), as measured by X-Ray powderdiffraction and dissolution rate by measurement of AUC.

In series of experiments with stable, amorphous hybrid nanoparticlescomprising nilotinib and HPMCP HP55 produced by the methods of theinvention, the resulting particles provided stable, amorphous hybridnanoparticles at 40% drug load (I/P nilotinib base/HPMCP HP55: exp 146),as measured by XRPD as well as dissolution rate by measurement of AUC.The material showed one glass transition temperature at ca 127° C.,which indicate a single amorphous phase with inherent stability.Partially crystalline batches also processed similar inherent stability.6 months storage at room temperature (18-25° C.) of partly crystallinehybrid nanoparticles at 40% drug load, UP nilotinib base/HPMCP HP55, didnot show any signs of physical instability. Thermalgravimetric analysisprovided a mass loss of 1.7% from ambient temperature to 120° C.

Dynamic vapor absorption analysis at 25° C. gave a relative massincrease of ca 7% from 0 to 90% RH (Three cycles from 0 to 90% RH, didnot induce a phase change).

The high glass transition temperature, 1.7% mass loss from ambienttemperature to 120° C. and moderate hygrospopicity propose an inherentstability. This is supported by stability testing of several batches atvarious conditions. The longest stability point is 12 months at roomtemperature (18-25° C.). No batches or conditions have shown any signsof physical instability (FIG. 27).

Modulated Differential Scanning Calorimetry (mDSC)

Modulated Differential Scanning calorimetry (mDSC) analysis was run on aTA Instruments Model Q200 (New Castle, USA), equipped with a RC90refrigerated cooling system (Home Automation, New Orleans, USA). Sampleswere weighed to 7±2 mg in Tzero Low-mass aluminum pans and sealed withTzero lids. They were then heated at a heating rate of 3° C./min from 0to 170° C. with conventional modulation temperature amplitude of 1° C.and a modulation period of 40 seconds. Ultra-high purity nitrogen wasused as purge gas at a flow rate of 50 mL/min. All data analyses wereperformed using TA Universal Analysis software, version 4.7A. Cellconstant and temperature calibrations were conducted with the use of anindium standard prior to instrument operation. DSC results wereevaluated in terms of both forward and reverse components of heat flow.

Thermogravimetry (TG) was performed on a Seiko TG/DTA 6200 and open 90μl Pt-pans with ca 10 to 20 mg of sample and a nitrogen flow of 200mL/min. The temperature program was ambient (20° C.) to 400° C. with aheating rate of 10° C./min. A blank was subtracted and the TG data wasnormalized with respect to sample size and analyzed using the MuseStandard Analysis software, version 6.1 U.

Dynamic Vapour Sorption (DVS)

The hygroscopicity of the samples was studied by Dynamic Vapor SorptionGravimetry (DVS), using a DVS-1 (Surface Measurement Ltd., UK).Approximately 10 mg of the substance was weighed into a glass cup. Therelative weight was recorded at 20 second interval when the targetrelative humidity (RH) over the sample was increased stepwise from 0% to90%, and then similarly decreased back to 0% RH, with 10% RH per step.Each sample was run in three consecutive full cycles. The condition toproceed to the next level of RH was a weight change below or equal to0.002% within 15 minutes, with a maximum total time per step of 24hours. Due to slow equilibration in experiments of this type, thenumbers obtained should be regarded as lower estimates of water uptake.The temperature was kept at 25° C.

X-Ray Powder Diffraction (XRPD)

XRD experiments were run on an X'Pert Pro diffractometer (PANanalytical,Almelo, Netherlands) set in Bragg-Brentano geometry. The diffractometerwas equipped with 20 μm nickel filter and an X'Celerator RTMS detectorwith an active length of 2.122° 2θ. A representative sample was placedon a zero background quarts single crystal specimen support (Siltronix,Archamps, France). Experiments were run using Cu K_(α) radiation (45 kVand 40 mA) at ambient temperature and humidity. Scans were run incontinuous mode in the range 4.5-40°2θ using automatic divergence andanti-scatter slits with observed length of 10 mm, a common counting timeof 299.72 seconds, and step size of 0.0167° 2θ. Data collection was doneusing the application software X'Pert Data Collector V.2.2j andinstrument control software V.2.1E, while pattern analysis was doneusing X'Pert Data Viewer V.1.2c (all software being from PANanalytical,Almelo, Netherlands).

Dissolution Rate by Measurement of AUC

The stable hybrid nanoparticles described in Exp 171 & Exp172 (I/P) asset out below, were produced according to Exp 148, with nilotinib base,HPMCP HP55 and stored at room temperature for 11 months. The non-sinkdissolution rate was tested at different different time points and theresults are presented in Table 47 and FIG. 26. Polyvinylcaprolactam-polyvinyl acetate-polyethylene glycol copolymer was added toenhance solubility. A comparison of the AUC over 80 minutes show clearlythat the dissolution rate profile of the particles is practicallyunchanged after 11 months storage, e.g. the ratio between the AUC ofparticles produced and tested, compared to particles produced, testedand stored for 11 months is over 97%.

TABLE 47 Ratio 0 1 min 5 min 10 min 20 min 40 min 80 min AUC (%) Exp 171(t = 0, 0 70.5 144.8 172.8 155.8 67.7 46.0 7411.9 n = 1) Exp 171 (t = 110 20.2 110.7 149.5 158.5 83.6 34.0 7234.5 97.6 months, n = 3) Standard 05.8 14.3 19.6 19.6 3.5 1.5 deviation

The invention claimed is:
 1. A pharmaceutical composition, comprising:(a) amorphous solid dispersion particles having a degree of amorphicityof 100% wherein the particles comprise (i) a protein kinase inhibitor inan amount of from about 10% by weight to about 70% by weight of theparticles; (ii) at least one polymeric stabilizing and matrix-formingcomponent; and (iii) at least one pharmaceutically acceptablesolubilizer selected from the group consisting of a d-α-tocopherol acidpolyethylene glycol 1000 succinate, a PEG-40 hydrogenated castor oil, aPEG-35 castor oil, a PEG-40 stearate, a hard fat, a polyoxylglyceride, aPEG-8 caprylic/capric glyceride, and a poloxamer; wherein the proteinkinase inhibitor is dasatinib, dasatinib hydrate, dasatinib solvate,dasatinib salt, or combinations thereof; and wherein the at least onepharmaceutically acceptable solubilizer is present in an amount of about0.5% by weight based on the weight of the particles and solubilizer. 2.The composition of claim 1, wherein the amount of the protein kinaseinhibitor is from about 10% by weight to about 50% by weight of theparticles.
 3. The composition of claim 1, wherein the amount of theprotein kinase inhibitor is from about 10% by weight to about 40% byweight of the particles.
 4. The composition of claim 1, wherein theamount of the protein kinase inhibitor is from about 10% by weight toabout 30% by weight of the particles.
 5. The composition of claim 1,wherein the amount of the protein kinase inhibitor is from about 30% byweight to about 40% by weight of the particles.
 6. The composition ofclaim 1, wherein the protein kinase inhibitor is dasatinib.
 7. Thecomposition of claim 1, wherein the protein kinase inhibitor isdasatinib hydrate.
 8. The composition of claim 1, wherein the proteinkinase inhibitor is dasatinib solvate.
 9. The composition of claim 1,wherein the protein kinase inhibitor is dasatinib salt.
 10. Thecomposition of claim 1, wherein the at least one polymeric stabilizingand matrix-forming component is selected from methyl cellulose,hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropylmethylcellulose, hydroxypropyl methylcellulose acetate succinate,hydroxypropyl methylcellulose phthalate, polyvinylpyrrolidone, polyvinylacetate phthalate, copolyvidone, crospovidone, methacrylic acid andethylacrylate copolymer, methacrylate acid and methyl methacrylatecopolymer, polyethylene glycol, DL lactide/glycolide copolymer, polyDL-lactide, cellulose acetate phthalate, carbomer homopolymer Type A,carbomer homopolymer Type B, aminoalkyl methacrylate copolymers, andpoloxamers.
 11. The composition of claim 1, wherein the at least onepolymeric stabilizing and matrix-forming component is selected from thegroup consisting of hydroxypropyl methylcellulose phthalate,hydroxypropyl cellulose, copolyvidone, hydroxypropyl methylcelluloseacetate succinate, polyvinyl acetate phthalate, cellulose acetatephthalate and polyvinylpyrrolidone.
 12. The composition of claim 1,wherein the composition further comprises the at least onepharmaceutically acceptable solubilizer.
 13. The composition of claim12, wherein the solubilizer is selected from the group consisting ofd-α-tocopherol acid polyethylene glycol 1000 succinate and ahydrogenated castor oil.
 14. The composition of claim 12, wherein thesolubilizer is distributed to the surface of the particles.
 15. Thecomposition of claim 1, wherein the amorphous solid dispersion comprisesa powder.
 16. The composition of claim 1, wherein the particles have anaverage particle diameter size of less than: (i) about 1000 nm, (ii)about 500 nm, or (iii) about 250 nm.
 17. A pharmaceutical composition,consisting of: (a) amorphous solid dispersion particles having a degreeof amorphicity of 100%, where the particles comprise (i) a proteinkinase inhibitor in an amount of from about 10% by weight to about 70%by weight of the particles; and (ii) copolyvidone; and (iii) at leastone pharmaceutically acceptable solubilizer selected from the groupconsisting of a d-α-tocopherol acid polyethylene glycol 1000 succinate,a PEG-40 hydrogenated castor oil, a PEG-35 castor oil, a PEG-40stearate, a hard fat, a polyoxylglyceride, a PEG-8 caprylic/capricglyceride, and a poloxamer and (b) an excipient; wherein the proteinkinase inhibitor is dasatinib, dasatinib hydrate, dasatinib solvate,dasatinib salt, or combinations thereof; wherein the at least onepharmaceutically acceptable solubilizer is present in an amount of about0.5% by weight based on the weight of the particles and solubilizer. 18.The pharmaceutical composition of claim 17, wherein the amount ofprotein kinase inhibitor, based on the total weight of particles, isfrom about 10% by weight to about 40% by weight.
 19. The pharmaceuticalcomposition of claim 17, wherein the amount of protein kinase inhibitor,based on the total weight of particles, is from about 10% by weight toabout 30% by weight.
 20. The pharmaceutical composition of claim 17,wherein the amount of protein kinase inhibitor, based on the totalweight of particles, is from about 30% by weight to about 40% by weight.21. The composition of claim 17, wherein the amorphous solid dispersioncomprises a powder.
 22. The pharmaceutical composition of claim 17,wherein the particles have an average particle size of less than: (i)about 1000 nm, (ii) about 500 nm, or (iii) about 250 nm.
 23. Thecomposition of claim 17, wherein the protein kinase inhibitor isdasatinib.
 24. The composition of claim 17, wherein the protein kinaseinhibitor is dasatinib hydrate.
 25. The composition of claim 17, whereinthe protein kinase inhibitor is dasatinib solvate.
 26. The compositionof claim 17, wherein the protein kinase inhibitor is dasatinib salt. 27.A method of treating proliferative disorder in a patient in needthereof, comprising administering a therapeutically effective amount ofthe pharmaceutical composition of claim 18.